You Know My Name (Look Up The Number)

What is your h-index on Twitter?

This thought crossed my mind yesterday when I saw a tweet that was tagged #academicinsults

It occurred to me that a Twitter account is a kind of micro-publishing platform. So what would “publication metrics” look like for Twitter? Twitter makes analytics available, so they can easily be crunched. The main metrics are impressions and engagements per tweet. As I understand it, impressions are the number of times your tweet is served up to people in their feed (boosted by retweets). Engagements are when somebody clicks on the tweet (either a link or to see the thread or whatever). In publication terms, impressions would equate to people downloading your paper and engagements mean that they did something with it, like cite it. This means that a “h-index” for engagements can be calculated with these data.

For those that don’t know, the h-index for a scientist means that he/she has h papers that have been cited h or more times. The Twitter version would be a tweeter that has h tweets that were engaged with h or more times. My data is shown here:

TwitterAnalyticsMy twitter h-index is currently 36. I have 36 tweets that have been engaged with 36 or more times.

So, this is a lot higher than my actual h-index, but obviously there are differences. Papers accrue citations as time goes by, but the information flow on Twitter is so fast that tweets don’t accumulate engagement over time. In that sense, the Twitter h-index is less sensitive to the time a user has been active on Twitter, versus the real h-index which is strongly affected by age of the scientist. Other differences include the fact that I have “published” thousands of tweets and only tens of papers. Also, whether or not more people read my tweets compared to my papers… This is not something I want to think too much about, but it would affect how many engagements it is possible to achieve.

The other thing I looked at was whether replying to somebody actually means more engagement. This would skew the Twitter h-index. I filtered tweets that started with an @ and found that this restricts who sees the tweet, but doesn’t necessarily mean more engagement. Replies make up a very small fraction of the h tweets.

I’ll leave it to somebody else to calculate the Impact Factor of Twitter. I suspect it is very low, given the sheer volume of tweets.

Please note this post is just for fun. Normal service will (probably) resume in the next post.

Edit: As pointed out in the comments this post is short on “Materials and Methods”. If you want to calculate your ownTwitter h-index, go here. When logged in to Twitter, the analytics page should present your data (it may take some time to populate this page after you first view it). A csv can be downloaded from the button on the top-right of the page. I imported this into IgorPro (as always) to generate the plots. The engagements data need to be sorted in descending order and then the h-index can be found by comparing the numbers with their ranked position.

The post title is from the quirky B-side to the Let It Be single by The Beatles.

Pay You Back In Time

A colleague once told me that they only review three papers per year and then refuse any further requests for reviewing. Her reasoning was as follows:

  • I publish one paper a year (on average)
  • This paper incurs three peer reviews
  • Therefore, I owe “the system” three reviews.

It’s difficult to fault this logic. However, I think that as a senior scientist with a wealth of experience, the system would benefit greatly from more of her input. Actually, I don’t think she sticks rigorously to this and I know that she is an Academic Editor at a journal so, in fact she contributes much more to the system than she was letting on.

I thought of this recently when – in the space of one week – I got three peer review requests, which I accepted. I began to wonder about my own debit and credit in the peer review system. I only have reliable data from 2010.

Reviews incurred as an author are in gold (re-reviews are in pale gold), reviews completed as a peer are in purple (re-reviews are in pale purple). They are plotted cumulatively and the difference – or the balance – is shown by the markers. So, I have been in a constant state of owing the system reviews and I’m in no position to be turning down review requests.

In my defence, I was for two years Section Editor at BMC Cell Biology which means that I contributed more to the system that the plot shows. Another thing is reviews incurred/completed as a grant applicant/referee. I haven’t factored those in, but I think this would take the balance down further. I also comment on colleagues papers and grant applications.

Thinking back, I’ve only ever turned down a handful of peer review requests. Reasons being either that the work was too far outside my area of expertise or that I had a conflict of interest. I’ve never cited a balance of zero as a reason for not reviewing and this analysis shows that I’m not in this category.

In case any Editors are reading this… I’m happy to review work in my area, but please remember I currently have three papers to review!

The post title comes from a demo recording by The Posies that can be found on the At Least, At Last compilation on Not Lame Recordings.

Strange Things – update

My post on the strange data underlying the new impact factor for eLife was read by many people. Thanks for the interest and for the comments and discussion that followed. I thought I should follow up on some of the issues raised in the post.

To recap:

  1. eLife received a 2013 Impact Factor despite only publishing 27 papers in the last three months of the census window. Other journals, such as Biology Open did not.
  2. There were spurious miscites to papers before eLife published any papers. I wondered whether this resulted in an early impact factor.
  3. The Web of Knowledge database has citations from articles in the past referring to future articles!

1. Why did eLife get an early Impact Factor? It turns out that there is something called a partial Impact Factor.  This is where an early Impact Factor is awarded to some journals in special cases. This is described here in a post at Scholarly Kitchen. Cell Reports also got an early Impact Factor and Nature Methods got one a few years ago (thanks to Daniel Evanko for tweeting about Nature Methods’ partial Impact Factor). The explanation is that if a journal is publishing papers that are attracting large numbers of citations it gets fast-tracked for an Impact Factor.

2. In a comment, Rafael Santos pointed out that the miscites were “from a 2013 eLife paper to an inexistent 2010 eLife paper, and another miscite from a 2013 PLoS Computational Biology paper to an inexistent 2011 eLife paper”. The post at Scholarly Kitchen confirms that citations are not double-checked or cleaned up at all by Thomson-Reuters. It occurred to me that journals looking to game their Impact Factor could alter the year for citations to papers in their own journal in order to inflate their Impact Factor. But no serious journal would do that – or would they?

3. This is still unexplained. If anybody has any ideas (other than time travel) please leave a comment.

Strange Things

I noticed something strange about the 2013 Impact Factor data for eLife.

Before I get onto the problem. I feel I need to point out that I dislike Impact Factors and think that their influence on science is corrosive. I am a DORA signatory and I try to uphold those principles. I admit that, in the past, I used to check the new Impact Factors when they were released, but no longer. This year, when the 2013 Impact Factors came out I didn’t bother to log on to take a look. A chance Twitter conversation with Manuel Théry (@ManuelTHERY) and Christophe Leterrier (@christlet) was my first encounter with the new numbers.

Huh? eLife has an Impact Factor?

For those that don’t know, the 2013 Impact Factor is worked out by counting the total number of 2013 cites to articles in a given journal that were published in 2011 and 2012. This number is divided by the number of “citable items” in that journal in 2011 and 2012.

Now, eLife launched in October 2012. So it seems unfair that it gets an Impact Factor since it only published papers for 12.5% of the window under scrutiny. Is this normal?

I looked up the 2013 Impact Factor for Biology Open, a Company of Biologists journal that launched in January 2012* and… it doesn’t have one! So why does eLife get an Impact Factor but Biology Open doesn’t?**

elife-JIFLooking at the numbers for eLife revealed that there were 230 citations in 2013 to eLife papers in 2011 and 2012. One of which was a mis-citation to an article in 2011. This article does not exist (the next column shows that there were no articles in 2011). My guess is that Thomson Reuters view this as the journal existing for 2011 and 2012, and therefore deserving of an Impact Factor. Presumably there are no mis-cites in the Biology Open record and it will only get an Impact Factor next year. Doesn’t this call into question the veracity of the database? I have found other errors in records previously (see here). I also find it difficult to believe that no-one checked this particular record given the profile of eLife.

elfie-citesPerhaps unsurprisingly, I couldn’t track down the rogue citation. I did look at the cites to eLife articles from all years in Web of Science, the Thomson Reuters database (which again showed that eLife only started publishing in Oct 2012). As described before there are spurious citations in the database. Josh Kaplan’s eLife paper on UNC13/Tomosyn managed to rack up 5 citations in 2004, some 9 years before it was published (in 2013)! This was along with nine other papers that somehow managed to be cited in 2004 before they were published. It’s concerning enough that these data are used for hiring, firing and funding decisions, but if the data are incomplete or incorrect this is even worse.

Summary: I’m sure the Impact Factor of eLife will rise as soon as it has a full window for measurement. This would actually be 2016 when the 2015 Impact Factors are released. The journal has made it clear in past editorials (and here) that it is not interested in an Impact Factor and won’t promote one if it is awarded. So, this issue makes no difference to the journal. I guess the moral of the story is: don’t take the Impact Factor at face value. But then we all knew that already. Didn’t we?

* For clarity, I should declare that we have published papers in eLife and Biology Open this year.

** The only other reason I can think of is that eLife was listed on PubMed right away, while Biology Open had to wait. This caused some controversy at the time. I can’t see why a PubMed listing should affect Impact Factor. Anyhow, I noticed that Biology Open got listed in PubMed by October 2012, so in the end it is comparable to eLife.

Edit: There is an update to this post here.

Edit 2: This post is the most popular on Quantixed. A screenshot of visitors’ search engine queries (Nov 2014)…

searches

The post title is taken from “Strange Things” from Big Black’s Atomizer LP released in 1986.

Vitamin K

Note: this is not a serious blog post.

Neil Hall’s think piece in Genome Biology on the Kardashian index (K-index) caused an online storm recently, spawning hashtags and outrage in not-so-equal measure. Despite all the vitriol that headed Neil’s way, very little of it concerned his use of Microsoft Excel to make his plot of Twitter followers vs total citations! Looking at the plot with the ellipse around a bunch of the points and also at the equations, I thought it might be worth double-checking Neil’s calculations.

In case you don’t know what this is about: the K-index is the ratio of actual Twitter followers (\(F_{a}\)) to the number of Twitter followers you are predicted to have (\(F_{c}\)) based on the total number of citations to your papers (\(C\)) from the equation:

\(F_{c}=43.3C^{0.32} \)

So the K-index is:

\(K-index=\frac{F_{a}}{F_{c}}\)

He argues that if a scientist has a K-index >5 then they are more famous for their twitterings than for their science. This was the most controversial aspect of the piece. It wasn’t clear whether he meant that highly cited scientists should get tweeting or that top-tweeters should try to generate some more citations (not as easy as it sounds). The equation for \(F_{c}\) was a bit suspect, derived from some kind of fit through some of the points. Anyway, it seemed to me that the ellipse containing the Kardashians didn’t look right.

I generated the data for \(F_{c}\) and for a line to show the threshold at which one becomes a Kardashian (k) in IgorPro as follows:

Make /o /N=100000 fc
fc =43.3*(x^0.32)
Duplicate fc k //yes, this does look rude
k *=5
display fc, k //and again!

K-index

This plot could be resized and overlaid on Neil’s Excel chart from Genome Biology. I kept the points but deleted the rest and then made this graph.

The Kardashians are in the peach zone. You’ll notice one poor chap is classed as a Kardashian by Neil, yet he is innocent! Clearly below the line, i.e. K-index <5.

Two confessions:

  1. My K-index today is 1.97 according to Twitter and Google Scholar.
  2. Embarrassingly, I didn’t know of the business person who gave her name to the K-index was until reading Neil’s article and the ensuing discussion. So I did learn something from this!

The post title is taken from “Vitamin K” by Gruff Rhys from the Hotel Shampoo album.

“Yeah” Is What We Had

When it comes to measuring the impact of our science, citations are pretty much all we have. And not only that but they only say one thing – yeah – with no context. How can we enrich citation data?

Much has been written about how and why and whether or not we should use metrics for research assessment. If we accept that metrics are here to stay in research assessment (of journals, Universities, departments and of individuals), I think we should be figuring out better ways to look at the available information.

8541947962_6853dd9786_zCitations to published articles are the key metric under discussion. This is because they are linked to research outputs (papers), have some relation to “impact” and they can be easily computed and a number of metrics have been developed to draw out information from the data (H-index, IF etc.). However there are many known problems with citations such as: they are heavily influenced by the size of the field. What I want to highlight here is what a data-poor resource they are and think of ways we could enrich the dataset with minimal modification to our existing databases.

1. We need a way to distinguish a yeah from a no

The biggest weakness of using citations as a measure of research impact is that a citation is a citation. It just says +1. We have no idea if +1 means “the paper stinks” or “the work is amazing!”.  It’s incredible that we can rate shoelaces on Amazon or eBay but we haven’t figured out a way to do this for scientific papers. Here’s a suggestion:

  • A neutral citation is +1
  • A positive citation is +2
  • A negative citation is -1

A neutral citation would be stating a fact and adding reference to support it, e.g. DNA is a double helix (Watson & Crick, 1953).

A positive citation would be something like: in agreement with Bloggs et al. (2010), we also find x.

A negative citation might be: we have tested the model proposed by Smith & Jones (1977) and find that it does not hold.

One further idea (described here) is to add more context to citation using keywords. Such as “replicating”, “using”, “consistent with”. This would also help with searching the scientific literature.

2. Multiple citations in one article

Because currently, citations are a +1, there is no way to distinguish whether the paper giving the citation was mentioning the cited paper in passing or was entirely focussed on that one paper.

Another way to think about this is that there are multiple reasons to cite a paper: maybe the method or reagent is being used, maybe they are talking about Figure 2 showing X or Figure 5 showing Y. What if a paper is talking about all of these things? In other words, the paper was very useful. Shouldn’t we record that interest?

Suggestion: A simple way to do this is to count the number of mentions in the text of the paper rather than just if the paper appears in the reference list.

3. Division of a citation unit for fair credit to each author

Calculations such as the H-index make no allowance for the position of the author in the author list (used in biological sciences and some other fields to denote contribution to the paper). It doesn’t make sense that the 25th author on a 50 author paper receives 100% of the citation credit as the first or last author. Similarly, the first author on a two author paper is only credited in the same way as the middle author on a multi-author paper. The difference in contribution is clear, but the citation credit is not. This is because the citation credit for the former paper is worth 25 times that of the latter! This needs to be equalised. The citation unit, c could be divided to achieve fair credit for authors. At the moment, c=1, but could be multiples (or negative values) as described above. Here’s a suggestion:

  • First (and multiple first) and last (and co-last) authors get 0.5c divided by number of authors.
  • The remainder, 0.5c, is divided between all authors.

For a two author paper: first author gets 0.5c and last author gets 0.5c. (0.5c/2+0.5c/2)=0.5c

For a ten author paper with one first author and one last author, first and last author each get (0.5c/2+0.5c/10)=0.3c and the 5th author gets (0c+0.5c/10)=0.05c.

Note that the sum for all authors will equal c. So this is equalised for all papers. These citation credits would then be the basis for H-index and other calculations for individuals.

Most simply, the denominator would be the number of authors, or – if we can figure out a numerical credit system – each author could be weighted according to their contribution.

4. Citations to reviews should be downgraded

A citation to a review is not equal to a citation to a research paper. For several reasons. First, they are cited at a higher rate, because they are a handy catchall citation particularly for the Introduction section in papers. This isn’t fair either and robs credit from the people who did the work that actually demonstrated what is being discussed. Second, the achievement of publishing a review is nothing in comparison to publishing a paper. Publishing a review involves 1) being asked, 2) writing it, 3) light peer review and some editing and that’s it! Publishing a research paper involves much more effort: having the idea, getting the money, hiring the people, training the people, getting a result – and we are only at the first panel in Fig 1A. Not to mention the people-hours and arduous peer review process. It’s not fair that citations to reviews are treated as equal to papers when it comes to research assessment.

Suggestion: a citation to a review should be worth a fraction (maybe 1/10th) of a citation to a research paper.

In addition, there are too many reviews written at the moment. I think this is not because they are particularly useful. Very few actually contribute a new view or new synthesis of an area, most are just a summary of the area. Journals like them because they drive up their citation metrics. Authors like them because it is nice to be invited to write something – it means people are interested in what you have to say… If citations to reviews were downgraded, there would be less incentive to publish them and we would have more space for all those real papers that are getting rejected at journals that claim that space is a limitation for publication.

5. Self-citations should be eliminated

If we are going to do all of the above, then self-citation would pretty soon become a problem. Excessive self-citation would be difficult to police, and not many scientists would go for a -1 citation to their own work. So, the simplest thing to do is to eliminate self-citation. Author identification is crucial here. At the moment this doesn’t work well. In ISI and Scopus, whatever algorithm they use keeps missing some papers of mine (and my name is not very common at all). I know people who have been grouped with other people that they have published one or two papers with. For authors with ambiguous names, this is a real problem. ORCID is a good solution and maybe having an ORCID (or similar) should be a requirement for publication in the future.

Suggestion: the company or body that collates citation information needs to accurately assign authors and make sure that research papers are properly segregated from reviews and other publication types.

These were five things I thought of to enrich citation data to improve research assessment, do you have any other ideas?

The post title is taken from ‘”Yeah” Is What We Had’ by Grandaddy from their album Sumday.

Round and Round

I thought I’d share a procedure for rotating a 2D set of coordinates about the origin. Why would you want do this? Well, we’ve been looking at cell migration in 2D – tracking nuclear position over time. Cells migrate at random and I previously blogged about ways to visualise these tracks more clearly. Part of this earlier procedure was to set the start of each track at (0,0). This gives a random hairball of tracks moving away from the origin. Wouldn’t it be a good idea to orient all the tracks so that the endpoint lies on the same axis? This would simplify the view and allow one to assess how ‘directional’ the cell tracks are. To rotate a set of coordinates, you need to use a rotation matrix. This allows you to convert the x,y coordinates to their new position x’,y’. This rotation is counter-clockwise.

\(x’ = x \cos \theta – y \sin \theta\,\)

\(y’ = x \sin \theta + y \cos \theta\,\)

However, we need to find theta first. To do this we need to find the angle between two lines, using this formula.

\(\cos \theta = \frac {\mathbf a \cdot \mathbf b}{\left \Vert {\mathbf a} \right \Vert \cdot \left \Vert {\mathbf b} \right \Vert} \)

The maths is kept to a minimum here. If you are interested, look at the code at the bottom.

beforeThe two lines (a and b) are formed by the x-axis (origin to some point on the x-axis, i.e. y=0) and by a line running from the origin to the last coordinate in the series. This calculation can be done for each track with theta for each track being used to rotate the that whole track (x,y changed to x’,y’ for each point).

Here is an example of just a few tracks from an experiment. Typically we have hundreds of tracks for each experimental group and the code will blast through them all very quickly (<1 s).

after

After rotation, the tracks are now aligned so that the last point is on the x-axis at y=0. This allows us to see how ‘directional’ the tracks are. The end points are now aligned, when they migrated there, how convoluted was their path.

The code to do this is up on Igor Exchange code snippets. A picture of the code is below (markup for code in WordPress is not very clear). See the code snippet if you want to use it.

rotator

The weakness of this method is that acos (arccos) only gives results from 0 to Pi (0 to 180°). There is a correction in the procedure, but everything needs editing if you want to rotate the co-ordinates to some other plane. Feedback welcome.

Edit Jim Prouty and A.G. have suggested two modifications to the code. The first is to use complex waves rather than 2D real waves. Then use two native Igor functions r2polar or p2rect. The second suggestion is to use Matrix operations! As is often the case with Igor there are several ways of doing things. The method described here is long-winded compared to a MatrixOp and if the waves were huge these solutions would be much, much faster. As it is, our migration movies typically have 60 points and as mentioned rotator() blasts through them very quickly. More complex coordinate sets would need something more sophisticated.

The post title is taken from “Round & Round” by New Order from their Technique LP.

Sure To Fall

What does the life cycle of a scientific paper look like?

It stands to reason that after a paper is published, people download and read the paper and then if it generates sufficient interest, it will begin to be cited. At some point these citations will peak and the interest will die away as the work gets superseded or the field moves on. So each paper has a useful lifespan. When does the average paper start to accumulate citations, when do they peak and when do they die away?

Citation behaviours are known to be very field-specific. So to narrow things down, I focussed on cell biology and in one area “clathrin-mediated endocytosis” in particular. It’s an area that I’ve published in – of course this stuff is driven by self-interest. I downloaded data for 1000 papers from Web of Science that had accumulated the most citations. Reviews were excluded, as I assume their citation patterns are different from primary literature. The idea was just to take a large sample of papers on a topic. The data are pretty good, but there are some errors (see below).

Number-crunching (feel free to skip this bit): I imported the data into IgorPro making a 1D wave for each record (paper). I deleted the last point corresponding to cites in 2014 (the year is not complete). I aligned all records so that year of publication was 0. Next, the citations were normalised to the maximum number achieved in the peak year. This allows us to look at the lifecycle in a sensible way. Next I took out records to papers less than 6 years old as I reasoned these would have not have completed their lifecycle and could contaminate the analysis (it turned out to make little difference). The lifecycles were plotted and averaged. I also wrote a quick function to pull out the peak year for citations post hoc.

So what did it show?

Citations to a paper go up and go down, as expected (top left). When cumulative citations are plotted most of the articles have an initial burst and then level off. The exception are ~8 articles that continue to rise linearly (top right). On average a paper generates its peak citations three years after publication (box plot). The fall after this peak period is pretty linear and it’s apparently all over somewhere >15 years after publication (bottom left). To look at the decline in more detail I aligned the papers so that year 0 was the year of peak citations. The average now loses almost 40% of those peak citations in the following year and then declines steadily (bottom right).

Edit: The dreaded Impact Factor calculation takes the citations to articles published in the preceding 2 years and divides by the number of citable items in that period. This means that each paper only contributes to the Impact Factor in years 1 and 2. This is before the average paper reaches its peak citation period. Thanks to David Stephens (@david_s_bristol) for pointing this out. The alternative 5 year Impact Factor gets around this limitation.

Perhaps lifecycle is the wrong term: papers in this dataset don’t actually ‘die’, i.e. go to 0 citations. There is always a chance that a paper will pick up the odd citation. Papers published 15 years ago are still clocking 20% of their peak citations. Looking at papers cited at lower rates would be informative here.

Two other weaknesses that affect precision is that 1) a year is a long time and 2) publication is subject to long lag times. The analysis would be improved by categorising the records based on the month-year when the paper was published and the month-year when each citation comes in. Papers published in January in one year probably have a different peak than those published in December of the same year, but this is lost when looking at year alone. Secondly, due to publication lag, it is impossible to know when the peak period of influence for a paper truly is.
MisCytesProblems in the dataset. Some reviews remained despite being supposedly excluded, i.e. they are not properly tagged in the database. Also, some records have citations from years before the article was published! The numbers of citations are small enough to not worry for this analysis, but it makes you wonder about how accurate the whole dataset is. I’ve written before about how complete citation data may or may not be. These sorts of things are a concern for all of us who are judged by these things for hiring and promotion decisions.

The post title is taken from ‘Sure To Fall’ by The Beatles, recorded during The Decca Sessions.

Tips from the Blog I

What is the best music to listen to while writing a manuscript or grant proposal? OK, I know that some people prefer silence and certainly most people hate radio chatter while trying to concentrate. However, if you like listening to music, setting an iPod on shuffle is no good since a track by Napalm Death can jump from the speakers and affect your concentration. Here is a strategy for a randomised music stream of the right mood and with no repetition, using iTunes.

For this you need:
A reasonably large and varied iTunes library that is properly tagged*.

1. Setup the first smart playlist to select all songs in your library that you like to listen to while writing. I do this by selecting genres that I find conducive to writing.
Conditions are:
-Match any of the following rules
-Genre contains jazz
-add as many genres as you like, e.g. shoegaze, space rock, dream pop etc.
-Don’t limit and do check live updating
I call this list Writing

2. Setup a second smart playlist that makes a randomised novel list from the first playlist
Conditions are:
-Match all of the following rules
-Playlist is Writing   //or whatever you called the 1st playlist
-Last played is not in the last 14 days    //this means once the track is played it disappears, i.e. refreshes constantly
-Limit to 50 items selected by random
-Check Live updating
I call this list Writing List

That’s it! Now play from Writing List while you write. The same strategy works for other moods, e.g. for making figures I like to listen to different music and so I have another pair for that.

After a while, the tracks that you’ve skipped (for whatever reason) clog up the playlist. Just select all and delete from the smart playlist, this refreshes the list and you can go again with a fresh set.

* If your library has only a few tracks, or has plenty of tracks but they are all of a similar genre, this tip is not for you.

Outer Limits

This post is about a paper that was recently published. It was the result of a nice collaboration between me and Francisco López-Murcia and Artur Llobet in Barcelona.

The paper in a nutshell
The availability of clathrin sets a limit for presynaptic function

Background
Clathrin is a three legged protein that forms a cage around membranes during endoctosis. One site of intense clathrin-mediated endocytosis (CME) is the presynaptic terminal. Here, synaptic vesicles need to be recaptured after fusion and CME is the main route of retrieval. Clathrin is highly abundant in all cells and it is generally thought of as limitless for the formation of multiple clathrin-coated structures. Is this really true? In a neuron where there is a lot of endocytic activity, maybe the limits are tested?
It is known that strong stimulation of neurons causes synaptic depression – a form of reversible synaptic plasticity where the neuron can only evoke a weak postsynaptic response afterwards. Is depression a vesicle supply problem?

What did we find?
We showed that clathrin availability drops during stimulation that evokes depression. The drop in availability is due to clathrin forming vesicles and moving away from the synapse. We mimicked this by RNAi, dropping the clathrin levels and looking at synaptic responses. We found that when the clathrin levels drop, synaptic responses become very small. We noticed that fewer vesicles are able to be formed and those that do form are smaller. Interestingly, the amount of neurotransmitter (acetylcholine) in the vesicles was much less than the volume of the vesicles as measured by electron microscopy. This suggests there is an additional sorting problem in cells with lower clathrin levels.

Killer experiment
A third reviewer was called in (due to a split decision between Reviewers 1 and 2). He/she asked a killer question: all of our data could be due to an off-target effect of RNAi, could we do a rescue experiment? We spent many weeks to get the rescue experiment to work, but a second viral infection was too much for the cells and engineering a virus to express clathrin was very difficult. The referee also said: if clathrin levels set a limit for synaptic function, why don’t you just express more clathrin? Well, we would if we could! But this gave us an idea… why don’t we just put clathrin in the pipette and let it diffuse out to the synapses and rescue the RNAi phenotype over time? We did it – and to our surprise – it worked! The neurons went from an inhibited state to wild-type function in about 20 min. We then realised we could use the same method on normal neurons to boost clathrin levels at the synapse and protect against synaptic depression. This also worked! These killer experiments were a great addition to the paper and are a good example of peer review improving the paper.

People
Fran and Artur did almost all the experimental work. I did a bit of molecular biology and clathrin purification. Artur and I wrote the paper and put the figures together – lots of skype and dropbox activity.
Artur is a physiologist and his lab like to tackle problems that are experimentally very challenging – work that my lab wouldn’t dare to do – he’s the perfect collaborator. I have known Artur for years. We were postdocs in the same lab at the LMB in the early 2000s. We tried a collaborative project to inhibit dynamin function in adrenal chromaffin cells at that time, but it didn’t work out. We have stayed in touch and this is our first paper together. The situation in Spain for scientific research is currently very bad and it deteriorated while the project was ongoing. This has been very sad to hear about, but fortunately we were able to finish this project and we hope to work together more in the future.

We were on the cover!
25.coverNow the scientific literature is online, this doesn’t mean so much anymore, but they picked our picture for the cover. It is a single cell microculture expressing GFP that was stained for synaptic markers and clathrin. I changed the channels around for artistic effect.

What else?
J Neurosci is slightly different to other journals that I’ve published in recently (my only other J Neurosci paper was published in 2002). For the following reasons:

  1. No supplementary information. The journal did away with this years ago to re-introduce some sanity in the peer review process. This didn’t affect our paper very much. We had a movie of clathrin movement that would have gone into the SI at another journal, but we simply removed it here.
  2. ORCIDs for authors are published with the paper. This gives the reader access to all your professional information and distinguishes authors with similar names. I think this is a good idea.
  3. Submission fee. All manuscripts are subject to a submission fee. I believe this is to defray the costs of editorial work. I think this makes sense, although I’m not sure how I would feel if our paper had been rejected.

Reference:

López-Murcia, F.J., Royle, S.J. & Llobet, A. (2014) Presynaptic clathrin levels are a limiting factor for synaptic transmission J. Neurosci., 34: 8618-8629. doi: 10.1523/JNEUROSCI.5081-13.2014

Pubmed | Paper

The post title is taken from “Outer Limits” a 7″ Single by Sleep ∞ Over released in 2010.