Proapoptotic Role of Human Growth and Transformation-Dependent Protein in the Developing Rat Brain After Hypoxia-Ischemia

Abstract

Background and Purpose-Human growth and transformation-dependent protein (HGTD-P) is a new proapoptotic protein and an effector of cell death induced by hypoxia-ischemia (HI). The function of HGTD-P has been investigated in human prostate cancer cells and mouse neurons cultured in vitro. However, whether HGTD-P is involved in regulating the apoptosis of rat neurons is not clear, and the relevance of HGTD-P in HI animal models is still unknown. Therefore, in the present study, we tried to elucidate the role that HGTD-P plays in apoptosis of rat neurons subjected to HI, both in culture and in the developing rat brain in vivo. Methods-Samples from primary cultured neurons and postnatal day 10 rat brains with HI were collected. RT-PCR, Western blotting, and immunocytochemistry were used to detect the expression and distribution of rat HGTD-P, cleaved caspase 3, and apoptosis-inducing factor (AIF). MTT assay, DAPI, TUNEL, and flowcytometry were used to detect cell viability and apoptosis. Results-We found that HI upregulated the mRNA and protein levels of HGTD-P in rat neurons in vitro and in vivo.Antisense oligonucleotides (AS) targeted to HGTD-P inhibited the expression of HGTD-P, thus rescuing neuronal viability and attenuating neuronal apoptosis. In addition, we found that HGTD-P played its proapoptotic role by activating caspase 3 and inducing the translocation of AIF to nuclear. Conclusions-Our findings show that HGTD-P plays a proapoptotic role in the developing rat brain after HI and that it may be a potential target in treating HI-induced brain damage.

Intestinal ischemia/reperfusion enhances microglial activation and induces cerebral injury and memory dysfunction in rats*

“…The protein expressions of cleaved caspase-3, a marker of apoptosis (24), and ionized calcium binding adaptor molecule 1 (Iba1) reflecting the activation of microglia (25) were detected by immunohistochemistry method. Briefly, the sections were incubated with rabbit anti-cleaved caspase-3 antibody (1:500; Cell Signaling Technology, Danvers, MA) and rabbit anti-Iba1 antibody (1:800; Wako Pure Chemicals, Osaka, Japan), respectively.…”

“…The procedure was performed as previously described (24). For cytokines detection, sections were incubated with goat polyclonal anti-rat TNF-α (1:50; Santa Cruz Biotechnology, Santa Cruz, CA) or goat polyclonal antirat IL-6 (1:50; SantaCruz Biotechnology).…”

“…The apoptosis of neural cell was detected with TUNEL method. Apoptotic cells were assessed using an assay kit (Roche, Indianapolis, IN) as previous study showed (24). For quantification, the mean number of TUNEL-positive cells in specific brain area was calculated.…”

Orphan proteins of unknown function in the mitochondrial intermembrane space proteome: New pathways and metabolic cross-talk

“…hFAM162A is speculated to be involved in hypoxia-induced cell death with cytochrome c release and caspase activation and inducing mitochondrial permeability transition. In neuronal cells, hFAM162A might promote release of hAIFM1 from mitochondria to the cytoplasm followed by its translocation to the nucleus; however, the involvement of caspases has been debated upon in the literature [80], [81]. The protein interacts with hHSP90AB, which is essential for hFAM162A mitochondrial localization and pro-apoptotic activity.…”

Sulforaphane protects primary cultures of cortical neurons against injury induced by oxygen-glucose deprivation/reoxygenation via antiapoptosis

“…The pathophysiology involves multiple biochemical mechanisms such as the generation of reactive oxygen species, glutamatemediated excitotoxicity, inflammation and apoptosis [1] . One of the central mechanisms of hypoxic-ischemic brain injury is neuronal loss caused by apoptosis [2,3] . The Bcl-2 family and caspases play key roles in the apoptotic cascade [4] .…”