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Abstract

This study set out to investigate, for the first time, the distribution and colocalization pattern of cocaine-and amphetamine-regulated transcript (CART) and one of the calcium binding-proteins: parvalbumin (PV) in the chinchilla’s hippocampus proper (HP). HP, consisting of Ammon’s horn (CA) and the dentate gyrus (DG), is an important component of the limbic system, involved in learning and memory processes. CA showed a higher immunoreactivity of CART (-IR) compared to DG. CART-IR neurons were mainly observed in the molecular layer of DG and in the pyramidal layer of CA. CART-IR fibers were present in the granular layer; in the hilus numerous mossy fibers were detected, while in the molecular layer CART-IR fibers were not found. In all CA fields (CA1-CA3), CART-IR fibers were only present in the lacuno- sum-molecular layer. Immunofluorescence with double- labeling showed that only CART-IR cells stained positive for PV, whereas in CART-IR fibers there was no PV-positive reaction. Our research supplements missing knowledge about the distribution and colocalization pattern of CART with PV in the chinchilla’s hippocampus, and also provides a better understanding of the similarities and differences among individuals of the same species and also with other mammals.

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Authors and Affiliations

R. Szalak
M. Matysek
W. Kukula-Koch
K. Rycerz
A. Zacharko-Siembida
M.B. Arciszewski
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Abstract

The process of vitrification of the cumulus-oocyte complex (COCs) often results in cold shock. When warming, heat shock occurs which can disrupt the balance of intracellular calcium (Ca2+) intensity. Drastic changes in temperature cause Reactive Oxygen Species (ROS), affecting changes on Ca2+ in COCs. The role of calcium is needed for oocyte activation in the fertilization process. The purpose of this study was to measure the expression of Ca2+ and the intensity of Ca2+ in COCs after vitrification. The study was divided into 2 groups, the control group (C) of fresh COCs, and the treatment group (T) of COCs after vitrification. After vitrification for 24 hours, then thawing, the expression of Ca2+ was examined using the Immunocytochemistry (ICC) method and the intensity of calcium (Ca2+) with a Confocal Laser Scanning Microscope (CLSM). The research data obtained were analyzed statistically by T-Test. The results showed that the expression of Ca2+ in the control group (12.00±0.00) was different from the treatment group (0.35±0.79). The intensity of Ca2+ in the control group (1059.43±489.59) was different from the treatment group (568.21±84.31). The conclusion of this study is that cryopreservation affects calcium in COCs; there were differences in the expression and the intensity of Ca2+ between fresh COCs and COCs after vitrification. Ca2+ intensity of COCs after vitrification was concentrated in the nucleus, while in fresh COCs it was concentrated in the cytoplasm.
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Authors and Affiliations

W. Widjiati
1
Z. Faizah
2
N. Darsini
2
V.F. Hendrawan
3
H.N. Karima
4
C. Chotimah
4
S.B. Sumitro
5
L.R. Yustinasari
6
A.A.M.N. Kasman
7
J.M. Ntoruru
8
E.M. Luqman
6

  1. Post Graduate School of Universitas Airlangga Surabaya, Indonesia
  2. Department of Biomedical Science, Faculty of Medicine Universitas Airlangga Surabaya, Indonesia
  3. Department of Reproduction, Faculty of Veterinary Medicine, Universitas Brawijaya Malang, Indonesia
  4. Bio-Science Central Laboratory, Universitas Brawijaya Malang, Indonesia
  5. Department of Biology, Faculty of Science, Universitas Brawijaya Malang, Indonesia
  6. Department of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga Surabaya, Indonesia
  7. Faculty of Health Science, Universitas Muhammadiyah Mataram, Mataram, Indonesia
  8. Research Assistant, Meru University, Kenya
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Abstract

Accumulation of LaCl3, a well-known Ca2+-channel blocker, can inhibit plant growth. However, the current understanding of its effects on gene expression is limited. In this paper, different concentrations of LaCl3 (0, 0.5, 1.0, 1.5, 2.0 mM) were used to treat germinated wheat ( Triticum aestivum L.) seeds for 24 h. The degree of root growth inhibition gradually increased with increasing LaCl3 concentration. qRT-PCR analysis revealed that the expression of several key genes related to the cell cycle process, such as pcna, mcm2, rdr and cyclin B, were significantly down-regulated. Further analysis of genomic DNA instability using Random Amplified Polymorphic DNA (RAPD) and methylation levels by Coupled Restriction Enzyme Digestion-Random Amplification (CRED-RA) analysis indicated a significant increase in genomic DNA polymorphisms and methylation levels. The results of this study verified that the reasons why LaCl3 treatment can inhibit the growth of wheat roots are as follows: interference in the normal progression of the cell cycle, induction of genomic DNA instability and increase in DNA methylation levels.
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Authors and Affiliations

Xia Lei
1
Keshi Ma
2
Feixiong Zhang
1

  1. College of Life Sciences, Capital Normal University, Beijing 100048, China
  2. College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou City, Henan Province, 466001, China

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