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README.md

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-# In-vivo_Apis2
 
-# Metadata for DOI registration according to DataCite Metadata Schema 4.1.
-# For detailed schema description see https://doi.org/10.5438/0014
-
-## Required fields
-
-# The main researchers involved. Include digital identifier (e.g., ORCID)
-# if possible, including the prefix to indicate its type.
 authors:
   -
     firstname: "Gérard"
@@ -28,16 +20,13 @@ authors:
     lastname: "Menzel"
     affiliation: "Freie Universität Berlin, Neurobiologie"
 
-
-# A title to describe the published resource.
 title: "In-vivo egfp expression in the honeybee Apis mellifera induced by electroporation and viral expression vector."
 
-# Additional information about the resource, e.g., a brief abstract.
-description: |
+description:
   In this study we describe egfp expression induced by two techniques: in vivo electroporation and viral transduction in several cell types of the adult honeybee brain. Non-neuronal and neuronal cell types were identified and the expression persisted at least during three days. Kenyon cells, optic lobe neurons and protocerebral lobe neurons were electroporated. Astrocyte-like glia cells, fibrous lamellar glia cells and cortex glia cells were identified. Viral transduction targeted one specific type of glia cells that could not be identified.  EGFP positive cells types were rather variable after electroporation, and viral transduction resulted in more homogenous groups of positive cells. We propose that these techniques remain a good alternative to transgenic animals because they potentially target only somatic cells.
 
-# Lit of keywords the resource should be associated with.
-# Give as many keywords as possible, to make the resource findable.
+Journal: PLoS ONE
+
 keywords:
   - Neuroscience
   - Insect
@@ -47,33 +36,15 @@ keywords:
   - viral transduction
   - eGFP
 
-# License information for this resource. Please provide the license name and/or a link to the license.
-# Please add also a corresponding LICENSE file to the repository.
-license:
-  name: "Creative Commons CC0 1.0 Public Domain Dedication"
-  url: "https://creativecommons.org/publicdomain/zero/1.0/"
+Fig 1. Specific egfp expression in the brain. B, D: Histological analyses of brain sections scanned at different wavelengths specific for native eGFP signals (GFP) or for the secondary antibody (CY5) or non-specific (CY3). In B, the plasmid was injected through the median ocellus and the electroporation was carried out with the large wire electrodes placed on the AL at 10 V with 3X 50 ms pulses. The animals were scarified 2 days after the electroporation. In D, the plasmid was injected through the median ocellus and the animals were electroporated with the large wire electrodes at 100 V with 5X 25ms pulses and sacrificed 1 day post-electroporation. The native eGFP and the CY5 signals overlay. D. Immunohistological analysis of eGFP signals 1 and 3 days post-electroporation. The plasmid was injected through the median ocellus and the electroporation was carried out with the plate electrodes at 100 V with 5X 50ms pulses (3 days post electroporation) and at 80 V with 5X 25ms pulses (1 day post electroporation). B, D and E: Section Z-axis 1 µm.
 
-## Optional Fields
+Fig 2. Distribution of anti-eGFP signals 3 days after different electroporation protocols. A: The animals were injected through the median ocellus and electroporated with the large wire electrodes. 100 V, 5X 25 ms. B: The animals were injected through the median ocellus and electroporated with the plate electrodes. The white arrowheads indicate perineural surface glia cells. 100 V, 5X 50 ms. C: The animals were injected into the MB calyx and electroporated with the plate electrodes. 100 V, 5X 25 ms. D: The animals were injected through the ocellus and electroporated with the small wire electrodes.100 V, 5X 25 ms. Section Z-axis 1 µM. 
 
-# Funding information for this resource.
-# Separate funder name and grant number by comma.
-funding:
-  - "DFG, LE1809/1-1"
-  - "DFG, LE1809/1-2"
+Fig 3. Non-neuronal transfected cell types. A: Anti-eGFP signals in cortex glia cells within the calycal region of the MB. The brain was electroporated 5X 25 ms pulses at 80 V with the small wire electrodes and the plasmid was injected in the ocellar tract. The brain was prepared 3 days post-electroporation. Section Z axis 0.5 µm. B: Anti-eGFP signal in cortex glia cells of the outer calycal region: The brain was electroporated with the large wire electrode 5X 25 ms pulses at 100 V and the plasmid was injected in the ocellar tract. Section Z axis 1 µm, C. Anti-eGFP signal in dendritic glia cells of the calyx: The brain was electroporated with the plate electrodes 5X 25 ms pulses at 100 V. Section Z axis 0.5 µm. D. Anti-eGFP signal in fibrous lamellar glia cells of the MB: The plasmid was injected directly into the MB and the brain was electroporated with plate electrodes 5X 25ms pulses at 100 V. Section Z axis 1 µm. The scans were performed at the Cy5 wavelength and in brightfield in alternance.
 
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-references:
-  -
-    id: "doi:tba"
-    reftype: "IsSupplementTo"
-    citation: "Citation1"
+Fig 4. Anti-eGFP signal detected in Kenyon cells: The animals were injected through the median ocellus and electroporated with the plate electrodes at 100 V, 5X 50 ms pulses. A. Section of the brain at the level of the suboesophageal ganglion (SEG). B. Section of the brain at the level of the central complex. The white arrow head shows the axons of KC terminating into the median lobe (Ml). eGFP positive signals were also detected into the calyx. C. Section of the brain at the level of the antennal lobe (AL). D. Section of the brain at the level of the antennal lobe (AL) and the vertical lobe (Vl).
+
+Fig 5. Anti eGFP signals detected in neuronal cell types: A. The plasmid was injected through the median ocellus. The brain was electroporated with the plate electrodes 5X 25 ms pulses at 100 V. Confocal sections (0.5 µm) B. The plasmid was injected through the median ocellus. The brain was electroporated with the plate electrodes 5X 25 ms pulses at 100 V. Section z-axis 1 µm. C. The plasmid was injected directly into the MB and the brain was electroporated 5X 25 ms pulses at 100 V with the plate electrodes. Section z-axis 1 µm. D. The brain was electroporated with the plate electrodes 5X 25 ms pulses at 100 V and the plasmid was injected directly into the MB. Section z-axis 1 µm. The scans were performed at the Cy5 wavelength and in brightfield in alternance.
 
-# Resource type. Default is Dataset, other possible values are Software, DataPaper, Image, Text.
-resourcetype: Dataset
+Fig 6. Anti-eGFP signals in bees injected with Lentiviruses. The bees were injected with viral particles through the median ocellus and the brain was dissected 3 days post-injection. Calyx (Ca), Suboesophageal ganglion (SEG).
 
-# Do not edit or remove the following line
-templateversion: 1.2