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Dissection of Drosophila Wandering Larval Brains for Patch-Clamping Neurons

Fernandez-Chiappe, Florencia ; Muraro, Nara I.

Cold Spring Harbor Laboratory ; 2022

In: Cold Spring Harbor Protocols Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107934-

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In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107934-

Abstract: An enormous amount of neuroscientific knowledge has been gained from studying the larval stage of Drosophila . From an electrophysiological point of view, the larval neuromuscular junction has played an important role in this quest for knowledge, as it presents practical advantages such as accessibility and a stereotypic pattern. The physiological properties of larval central neurons have been less explored, with information regarding mainly a few identified motoneurons available to date. This protocol describes a quick and easy dissection of the brain of wandering third-instar Drosophila larvae to produce an ex vivo preparation in which central neurons can be patch-clamped. Immobilizing fresh and tiny tissues, such as larval brains, to perform successful patch-clamp recordings is a crucial step; here we explain in detail how this can be achieved using cyanoacrylate glue.

Type of Medium: Online Resource

ISSN: 1940-3402 , 1559-6095

URL: Article

DOI: 10.1101/pdb.prot107934

Language: English

Publisher: Cold Spring Harbor Laboratory

Publication Date: 2022

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Patch-Clamping Drosophila Brain Neurons

Fernandez-Chiappe, Florencia ; Muraro, Nara I.

Cold Spring Harbor Laboratory ; 2022

In: Cold Spring Harbor Protocols Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107936-

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In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107936-

Abstract: Drosophila melanogaster is widely used as a model organism in all fields of biomedical research. In neuroscience, vast amounts of information have been gained using this little fly including the identification of neuronal circuits that regulate behaviors, the unraveling of their genetic underpinnings, and the molecular mechanisms involved. With plenty of genetic tools available to manipulate and infer neuronal activity, the direct measurement of electrical properties of fly neurons has lagged behind. This is due to the intricacies of performing electrical recordings in small cells such as fly central neurons. The patch-clamp technique offers the unique possibility of directly measuring the electrical properties of Drosophila neurons. This step-by-step protocol provides detailed advice for mastering this technique.

Type of Medium: Online Resource

ISSN: 1940-3402 , 1559-6095

URL: Article

DOI: 10.1101/pdb.prot107936

Language: English

Publisher: Cold Spring Harbor Laboratory

Publication Date: 2022

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Recommendations for empowering early career researchers to improve research culture and practice

Kent, Brianne A. ; Holman, Constance ; Amoako, Emmanuella ; [et al.]

Public Library of Science (PLoS) ; 2022

In: PLOS Biology Vol. 20, No. 7 ( 2022-7-7), p. e3001680-

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In: PLOS Biology, Public Library of Science (PLoS), Vol. 20, No. 7 ( 2022-7-7), p. e3001680-

Abstract: Early career researchers (ECRs) are important stakeholders leading efforts to catalyze systemic change in research culture and practice. Here, we summarize the outputs from a virtual unconventional conference (unconference), which brought together 54 invited experts from 20 countries with extensive experience in ECR initiatives designed to improve the culture and practice of science. Together, we drafted 2 sets of recommendations for (1) ECRs directly involved in initiatives or activities to change research culture and practice; and (2) stakeholders who wish to support ECRs in these efforts. Importantly, these points apply to ECRs working to promote change on a systemic level, not only those improving aspects of their own work. In both sets of recommendations, we underline the importance of incentivizing and providing time and resources for systems-level science improvement activities, including ECRs in organizational decision-making processes, and working to dismantle structural barriers to participation for marginalized groups. We further highlight obstacles that ECRs face when working to promote reform, as well as proposed solutions and examples of current best practices. The abstract and recommendations for stakeholders are available in Dutch, German, Greek (abstract only), Italian, Japanese, Polish, Portuguese, Spanish, and Serbian.

Type of Medium: Online Resource

ISSN: 1545-7885

URL: Article

DOI: 10.1371/journal.pbio.3001680

DOI: 10.1371/journal.pbio.3001680.g001

DOI: 10.1371/journal.pbio.3001680.g002

DOI: 10.1371/journal.pbio.3001680.t001

DOI: 10.1371/journal.pbio.3001680.s001

DOI: 10.1371/journal.pbio.3001680.s002

DOI: 10.1371/journal.pbio.3001680.s003

DOI: 10.1371/journal.pbio.3001680.s004

DOI: 10.1371/journal.pbio.3001680.s005

DOI: 10.1371/journal.pbio.3001680.s006

DOI: 10.1371/journal.pbio.3001680.s007

DOI: 10.1371/journal.pbio.3001680.s008

DOI: 10.1371/journal.pbio.3001680.s009

DOI: 10.1371/journal.pbio.3001680.s010

DOI: 10.1371/journal.pbio.3001680.s011

DOI: 10.1371/journal.pbio.3001680.s012

DOI: 10.1371/journal.pbio.3001680.s013

DOI: 10.1371/journal.pbio.3001680.s014

DOI: 10.1371/journal.pbio.3001680.s015

DOI: 10.1371/journal.pbio.3001680.s016

DOI: 10.1371/journal.pbio.3001680.s017

DOI: 10.1371/journal.pbio.3001680.s018

DOI: 10.1371/journal.pbio.3001680.s019

Language: English

Publisher: Public Library of Science (PLoS)

Publication Date: 2022

detail.hit.zdb_id: 2126773-X

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Ways to increase equity, diversity and inclusion

Mehta, Devang ; Bediako, Yaw ; de Winde, Charlotte M ; [et al.]

eLife Sciences Publications, Ltd ; 2020

In: eLife Vol. 9 ( 2020-07-07)

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In: eLife, eLife Sciences Publications, Ltd, Vol. 9 ( 2020-07-07)

Abstract: The eLife Early-Career Advisory Group (ECAG), an international group of early-career researchers committed to improving research culture, calls for radical changes at eLife and other journals to address racism in the scientific community and to make science more diverse and inclusive.

Type of Medium: Online Resource

ISSN: 2050-084X

URL: Article

DOI: 10.7554/eLife.60438

Language: English

Publisher: eLife Sciences Publications, Ltd

Publication Date: 2020

detail.hit.zdb_id: 2687154-3

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Effects of Pubertal Status and Inflammation on the Use of Ferritin to Define Iron Deficiency in Children With Overweight or Obesity

Muzzio, Maria Luz ; Lozano Chiappe, Ezequiel S ; Kabakian, Laura ; [et al.]

SAGE Publications ; 2019

In: Nutrition and Metabolic Insights Vol. 12 ( 2019-01), p. 117863881983906-

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In: Nutrition and Metabolic Insights, SAGE Publications, Vol. 12 ( 2019-01), p. 117863881983906-

Abstract: A worldwide increase in childhood overweight (OW) and obesity (OB) has been reported. OB is an inflammatory state which affects iron metabolism and the sensibility of the tests to detect iron deficiency (ID). Our aim was to evaluate the adequacy of current ferritin cut-offs to define ID in children with OW/OB. Methods: This cross-sectional study included 152 children (54% girls) aged (median [Q1-Q3]) 11 (8-13) years with OW/OB. Complete blood count and iron metabolism were evaluated. Low ferritin, transferrin saturation (TSat), and anemia were defined by age- and sex-specific cut-offs recommended by National Guidelines. Iron intake was assessed in a subgroup (n = 80) by a 24-hour dietary recall. Analyses were made according to pubertal development and ferritin tertiles. Results: The overall prevalence of low ferritin, TSat, and anemia was 2.6%, 23.8%, and 5.2%, respectively. Among pre-pubertal children (n = 87), the frequency of low TSat rose across ferritin tertiles ( P 〈 .05), whereas it decreased among pubertal children (n = 65; P 〈 .005). Cases of anemia among pre-pubertal children were found in the highest ferritin tertile, whereas 4/6 anemia cases in pubertal children were found in the lowest ferritin tertile ( 〈 39 µg/L). Pubertal children within the lowest ferritin tertile + low TSat (n = 11) showed lower hemoglobin (–9%; P 〈 .005) and hematocrit (–8%, P 〈 .01) than those in the same tertile + normal TSat (n = 16). The overall prevalence of children with ferritin 〈 39 µg/L + low TSat was 9.2%. Conclusions: Higher ferritin cut-off values are required to define ID in children with OW/OB. Such cut-off remains to be validated in larger, multi-ethnic cohorts of children with OW/OB.

Type of Medium: Online Resource

ISSN: 1178-6388 , 1178-6388

URL: Article

DOI: 10.1177/1178638819839064

Language: English

Publisher: SAGE Publications

Publication Date: 2019

detail.hit.zdb_id: 2594646-8

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Preparation of Pipettes and Pipette-Filling Devices for Patch-Clamping Drosophila Neurons

Fernandez-Chiappe, Florencia ; Muraro, Nara I.

Cold Spring Harbor Laboratory ; 2022

In: Cold Spring Harbor Protocols Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107932-

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In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2022, No. 8 ( 2022-08), p. pdb.prot107932-

Abstract: An essential requirement of every laboratory procedure is to have all materials ready when they are needed, so that the experimental flow is not disrupted. This is particularly true for patch clamping; therefore, effort must be devoted in advance to produce materials such as patch pipettes. This can be a fiddly business; hence, this protocol provides step-by-step advice on how to pull and polish patch-clamp pipettes. It also includes a brief description on how to prepare homemade filling devices to deliver saline efficiently and inexpensively into the pipettes. The protocol ends with guidelines on how to change the filament of a Sutter horizontal puller, a dreaded yet necessary activity that should be learned by anyone who wishes to become an expert patch clamper.

Type of Medium: Online Resource

ISSN: 1940-3402 , 1559-6095

URL: Article

DOI: 10.1101/pdb.prot107932

Language: English

Publisher: Cold Spring Harbor Laboratory

Publication Date: 2022

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Patch-Clamping Fly Brain Neurons

Fernandez-Chiappe, Florencia ; Muraro, Nara I.

Cold Spring Harbor Laboratory ; 2022

In: Cold Spring Harbor Protocols Vol. 2022, No. 8 ( 2022-08), p. pdb.top107796-

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In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2022, No. 8 ( 2022-08), p. pdb.top107796-

Abstract: The membrane potential of excitable cells, such as neurons and muscle cells, experiences a rich repertoire of dynamic changes mediated by an array of ligand- and voltage-gated ion channels. Central neurons, in particular, are fantastic computators of information, sensing, and integrating multiple subthreshold currents mediated by synaptic inputs and translating them into action potential patterns. Electrophysiology comprises a group of techniques that allow the direct measurement of electrical signals. There are many different electrophysiological approaches, but, because Drosophila neurons are small, the whole-cell patch-clamp technique is the only applicable method for recording electrical signals from individual central neurons. Here, we provide background on patch-clamp electrophysiology in Drosophila and introduce protocols for dissecting larval and adult brains, as well as for achieving whole-cell patch-clamp recordings of identified neuronal types. Patch clamping is a labor-intensive technique that requires a great deal of practice to become an expert; therefore, a steep learning curve should be anticipated. However, the instant gratification of neuronal spiking is an experience that we wish to share and disseminate, as many more Drosophila patch clampers are needed to study the electrical features of so many fly neuronal types unknown to date.

Type of Medium: Online Resource

ISSN: 1940-3402 , 1559-6095

URL: Article

DOI: 10.1101/pdb.top107796

Language: English

Publisher: Cold Spring Harbor Laboratory

Publication Date: 2022

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Mitigating the impact of conference and travel cancellations on researchers’ futures

Weissgerber, Tracey ; Bediako, Yaw ; de Winde, Charlotte M ; [et al.]

eLife Sciences Publications, Ltd ; 2020

In: eLife Vol. 9 ( 2020-03-27)

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In: eLife, eLife Sciences Publications, Ltd, Vol. 9 ( 2020-03-27)

Abstract: The need to protect public health during the current COVID-19 pandemic has necessitated conference cancellations on an unprecedented scale. As the scientific community adapts to new working conditions, it is important to recognize that some of our actions may disproportionately affect early-career researchers and scientists from countries with limited research funding. We encourage all conference organizers, funders and institutions who are able to do so to consider how they can mitigate the unintended consequences of conference and travel cancellations and we provide seven recommendations for how this could be achieved. The proposed solutions may also offer long-term benefits for those who normally cannot attend conferences, and thus lead to a more equitable future for generations of researchers.

Type of Medium: Online Resource

ISSN: 2050-084X

URL: Article

DOI: 10.7554/eLife.57032

Language: English

Publisher: eLife Sciences Publications, Ltd

Publication Date: 2020

detail.hit.zdb_id: 2687154-3

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eLife’s new model and its impact on science communication

Urban, Lara ; De Niz, Mariana ; Fernández-Chiappe, Florencia ; [et al.]

eLife Sciences Publications, Ltd ; 2022

In: eLife Vol. 11 ( 2022-12-08)

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In: eLife, eLife Sciences Publications, Ltd, Vol. 11 ( 2022-12-08)

Abstract: The eLife Early-Career Advisory Group discusses eLife’s new peer review and publishing model, and how the whole process of scientific communication could be improved for the benefit of early-career researchers and the entire scientific community.

Type of Medium: Online Resource

ISSN: 2050-084X

URL: Article

DOI: 10.7554/eLife.84816

Language: English

Publisher: eLife Sciences Publications, Ltd

Publication Date: 2022

detail.hit.zdb_id: 2687154-3

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High-Frequency Neuronal Bursting is Essential for Circadian and Sleep Behaviors in Drosophila

Fernandez-Chiappe, Florencia ; Frenkel, Lia ; Colque, Carina Celeste ; [et al.]

Society for Neuroscience ; 2021

In: The Journal of Neuroscience Vol. 41, No. 4 ( 2021-01-27), p. 689-710

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In: The Journal of Neuroscience, Society for Neuroscience, Vol. 41, No. 4 ( 2021-01-27), p. 689-710

Abstract: Circadian rhythms have been extensively studied in Drosophila ; however, still little is known about how the electrical properties of clock neurons are specified. We have performed a behavioral genetic screen through the downregulation of candidate ion channels in the lateral ventral neurons (LNvs) and show that the hyperpolarization-activated cation current I h is important for the behaviors that the LNvs influence: temporal organization of locomotor activity, analyzed in males, and sleep, analyzed in females. Using whole-cell patch clamp electrophysiology we demonstrate that small LNvs (sLNvs) are bursting neurons, and that I h is necessary to achieve the high-frequency bursting firing pattern characteristic of both types of LNvs in females. Since firing in bursts has been associated to neuropeptide release, we hypothesized that I h would be important for LNvs communication. Indeed, herein we demonstrate that I h is fundamental for the recruitment of pigment dispersing factor (PDF) filled dense core vesicles (DCVs) to the terminals at the dorsal protocerebrum and for their timed release, and hence for the temporal coordination of circadian behaviors. SIGNIFICANCE STATEMENT Ion channels are transmembrane proteins with selective permeability to specific charged particles. The rich repertoire of parameters that may gate their opening state, such as voltage-sensitivity, modulation by second messengers and specific kinetics, make this protein family a determinant of neuronal identity. Ion channel structure is evolutionary conserved between vertebrates and invertebrates, making any discovery easily translatable. Through a screen to uncover ion channels with roles in circadian rhythms, we have identified the I h channel as an important player in a subset of clock neurons of the fruit fly. We show that lateral ventral neurons (LNvs) need I h to fire action potentials in a high-frequency bursting mode and that this is important for peptide transport and the control of behavior.

Type of Medium: Online Resource

ISSN: 0270-6474 , 1529-2401

URL: Article

DOI: 10.1523/JNEUROSCI.2322-20.2020

Language: English

Publisher: Society for Neuroscience

Publication Date: 2021

detail.hit.zdb_id: 1475274-8

SSG: 12

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