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1.
Sci Rep ; 11(1): 24212, 2021 12 20.
Article in English | MEDLINE | ID: mdl-34930955

ABSTRACT

Fiber photometry (FP) is an adaptable method for recording in vivo neural activity in freely behaving animals. It has become a popular tool in neuroscience due to its ease of use, low cost, the ability to combine FP with freely moving behavior, among other advantages. However, analysis of FP data can be challenging for new users, especially those with a limited programming background. Here, we present Guided Photometry Analysis in Python (GuPPy), a free and open-source FP analysis tool. GuPPy is designed to operate across computing platforms and can accept data from a variety of FP data acquisition systems. The program presents users with a set of graphic user interfaces (GUIs) to load data and provide input parameters. Graphs are produced that can be easily exported for integration into scientific figures. As an open-source tool, GuPPy can be modified by users with knowledge of Python to fit their specific needs.


Subject(s)
Image Processing, Computer-Assisted/methods , Neuroimaging/instrumentation , Neuroimaging/methods , Photometry/instrumentation , Photometry/methods , Software , Algorithms , Animals , Area Under Curve , Artifacts , Brain/diagnostic imaging , Calcium/chemistry , Computer Graphics , Dopamine/chemistry , Female , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Neurosciences , Programming Languages , Stereotaxic Techniques , User-Computer Interface
2.
Gen Comp Endocrinol ; 222: 158-66, 2015 Oct 01.
Article in English | MEDLINE | ID: mdl-26141148

ABSTRACT

Secretoneurin (SN) in the preoptic area and pituitary of mammals and fish has a conserved close association with the vasopressin and oxytocin systems, members of a peptide family that are key in the modulation of sexual and social behaviors. Here we show the presence of SN-immunoreactive cells and projections in the brain of the electric fish, Brachyhypopomus gauderio. Secretoneurin colocalized with vasotocin (AVT) and isotocin in cells and fibers of the preoptic area. In the rostral pars distalis of the pituitary, many cells were both SN and prolactin-positive. In the hindbrain, at the level of the command nucleus of the electric behavior (pacemaker nucleus; PN), some of SN-positive fibers colocalized with AVT. We also explored the potential neuromodulatory role of SN on electric behavior, specifically on the rate of the electric organ discharge (EOD) that signals arousal, dominance and subordinate status. Each EOD is triggered by the command discharge of the PN, ultimately responsible for the basal EOD rate. SN modulated diurnal basal EOD rate in freely swimming fish in a context-dependent manner; determined by the initial value of EOD rate. In brainstem slices, SN partially mimicked the in vivo behavioral effects acting on PN firing rate. Taken together, our results suggest that SN may regulate electric behavior, and that its effect on EOD rate may be explained by direct action of SN at the PN level through either neuroendocrine and/or endocrine mechanisms.


Subject(s)
Electric Fish/genetics , Neuropeptides/metabolism , Secretogranin II/metabolism , Vasotocin/metabolism , Animals
3.
Neuroendocrinology ; 98(3): 212-23, 2013.
Article in English | MEDLINE | ID: mdl-24080944

ABSTRACT

BACKGROUND/AIMS: Adult mice lacking functional GABAB receptors (GABAB1KO) show altered Gnrh1 and Gad1 expressions in the preoptic area-anterior hypothalamus (POA-AH) and females display disruption of cyclicity and fertility. Here we addressed whether sexual differentiation of the brain and the proper wiring of the GnRH and kisspeptin systems were already disturbed in postnatal day 4 (PND4) GABAB1KO mice. METHODS: PND4 wild-type (WT) and GABAB1KO mice of both sexes were sacrificed; tissues were collected to determine mRNA expression (qPCR), amino acids (HPLC), and hormones (RIA and/or IHC). RESULTS: GnRH neuron number (IHC) did not differ among groups in olfactory bulbs or OVLT-POA. Gnrh1 mRNA (qPCR) in POA-AH was similar among groups. Gnrh1 mRNA in medial basal hypothalamus (MBH) was similar in WTs but was increased in GABAB1KO females compared to GABAB1KO males. Hypothalamic GnRH (RIA) was sexually different in WTs (males > females), but this sex difference was lost in GABAB1KOs; the same pattern was observed when analyzing only the MBH, but not in the POA-AH. Arcuate nucleus Kiss1 mRNA (micropunch-qPCR) was higher in WT females than in WT males and GABAB1KO females. Gad1 mRNA in MBH was increased in GABAB1KO females compared to GABAB1KO males. Serum LH and gonadal estradiol content were also increased in GABAB1KOs. CONCLUSION: We demonstrate that GABABRs participate in the sexual differentiation of the ARC/MBH, because sex differences in several reproductive genes, such as Gad1, Kiss1 and Gnrh1, are critically disturbed in GABAB1KO mice at PND4, probably altering the organization and development of neural circuits governing the reproductive axis.


Subject(s)
Glutamate Decarboxylase/deficiency , Gonadotropin-Releasing Hormone/deficiency , Hypothalamus, Middle/metabolism , Kisspeptins/deficiency , Protein Precursors/deficiency , Receptors, GABA-B/deficiency , Sex Differentiation/genetics , Animals , Animals, Newborn , Arcuate Nucleus of Hypothalamus/growth & development , Arcuate Nucleus of Hypothalamus/metabolism , Female , Gene Expression Regulation, Developmental , Glutamate Decarboxylase/genetics , Gonadotropin-Releasing Hormone/genetics , Hypothalamus, Middle/growth & development , Kisspeptins/genetics , Male , Mice , Mice, Inbred BALB C , Mice, Knockout , Protein Precursors/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Receptors, GABA-B/genetics
4.
Chemosphere ; 76(3): 374-80, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19394674

ABSTRACT

This study assessed the effects of cypermethrin and temperature on the survival, growth, sex differentiation, and gonadal development of Odontesthes bonariensis, a gonochoristic teleost with a strong thermolabile sex differentiation. Two complementary trials were conducted. In the first trial, newly hatched larvae were exposed during six weeks to 0 or 0.1 microgL(-1) of cypermethrin at 17, 22, and 29 degrees C. In the second trial, larvae were exposed at 22 degrees C to 0, 0.1, or 0.125 microgL(-1) of cypermethrin, or 100 microgL(-1) of the non-steroidal antiestrogen tamoxifen. Survival and growth of fish were affected by cypermethrin exposure, water temperature, and the combination of both factors. The survival rate decreased at higher temperatures and cypermethrin concentrations, but the insecticide lethality was inversely related to temperature. Growth was lower at 17 degrees C than at 22 or 29 degrees C, and was significantly increased by cypermethrin exposure. As already described for this species, all females or all males were obtained at 17 or 29 degrees C, respectively, and neither cypermethrin nor tamoxifen exposure caused changes in sex ratios. Slight changes in gonadal development were induced only by temperature. Finally, results showed that the in vitro antiestrogenic effect reported for cypermethrin had no in vivo effects on the sex ratio, the gonadal development, or the germ cell production of O. bonariensis, even at concentrations that affected the growth and survival of the fish.


Subject(s)
Environmental Exposure , Gonads/drug effects , Insecticides/toxicity , Pyrethrins/toxicity , Sex Differentiation/drug effects , Smegmamorpha/growth & development , Animals , Female , Gonads/growth & development , Larva/drug effects , Larva/growth & development , Male , Sex Ratio , Survival Rate , Temperature
5.
Gen Comp Endocrinol ; 143(1): 21-32, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15993101

ABSTRACT

The brain-type aromatase (CYP19A2) cDNA from pejerrey Odontesthes bonariensis was characterized. Its sequence differs from the ovarian-derived aromatase (CYP19A1) previously reported for the same species. The cDNA is 2305bp in length and the deduced protein comprises 501 amino-acids. The percentage of identity was higher when compared to other brain-derived aromatase proteins (85-63%) and lower with ovarian-derived aromatases (64-57%). Pejerrey aromatases share 61% of identity. The tissue expression analysis showed that CYP19A2 was expressed in the kidney, brain, and pituitary gland of both sexes and also in the ovary, but not in the eye, spleen, liver, gill, and testis. Semi-quantitative RT-PCR analysis of different brain areas revealed that CYP19A2 was expressed significantly higher in anterior male brain areas than in the corresponding female areas, and also when compared to posterior brain areas from both sexes. An immunological analysis using a polyclonal anti-teleost aromatase showed immunoreactive aromatase cells bordering the telencephalic ventricle and a strong signal in the ependymal cells of the preoptic area and the hypothalamus. In the optic tectum immunoreactive aromatase cells were labeled in the ventral wall and in the ependymal layer of the third and fourth ventricle with lateral projections. In the pituitary gland immunoreactive aromatase cells could be found in the rostral and proximal pars distalis. In this gland, aromatase fibers were also detected in different areas; many of them concentrated around blood vessels.


Subject(s)
Aromatase/metabolism , Brain/enzymology , DNA, Complementary , Fishes , Gene Expression Regulation, Enzymologic , Zebrafish Proteins/metabolism , Amino Acid Sequence , Animals , Base Sequence , DNA, Complementary/analysis , Female , Immunochemistry , Male , Molecular Sequence Data , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
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