Hidekatsu Mizushima

PACAP protects hippocampal neurons against apoptosis: Involvement of JNK/SAPK signaling pathway

Abstract: We have demonstrated that the ischemia‐induced apoptosis of neurons in the CA1 region of the rat hippocampus was prevented by either intracerebroventricular or intravenous infusion of pituitary adenylate cyclase‐activating polypeptide (PACAP). However, the molecular mechanisms underlying the anti‐apoptotic effect of PACAP remain to be determined. Within 3–6 h after ischemia, the activities of members of the mitogen‐activated protein (MAP) kinase family, including extracellular signal‐regulated kinase (ERK), Jun N‐terminal kinase (JNK)/stress‐activated protein kinase (SAPK), and p38 were increased in the hippocampus. The ischemic stress had a potent influence on the MAP kinase family, especially on JNK/SAPK. PACAP inhibited the activation of JNK/SAPK after ischemic stress. Secretion of interleukin‐6 (IL‐6) into the cerebrospinal fluid was intensely stimulated after PACAP infusion. IL‐6 inhibited the activation of JNK/SAPK, while it activated ERK. These observations suggest that PACAP and IL‐6 act to inhibit the JNK/SAPK signaling pathway, thereby protecting neurons against apoptosis.

Delayed neuronal cell death in the rat hippocampus is mediated by the mitogen-activated protein kinase signal transduction pathway

Transient global ischemia caused by 5 min of cardiac arrest induced delayed neuronal cell death (apoptosis) in the CA1 region of the rat hippocampus. To characterize the molecular mechanisms that regulate apoptosis in vivo, the contributions to cell death of mitogen-activated protein kinase family members were examined in the hippocampal region after brain ischemia-reperfusion. Ischemia-reperfusion led to a strong activation of the JNK/SAPK (c-Jun NH2-terminal protein kinase/stress activated protein kinase), ERK (extracellular signal-regulated kinase), and p38 enzymes. These results with other previous studies suggest that the activation of JNK/SAPK in accordance with p38 contributes to the induction of apoptosis in CA1 neurons.

Reduced postischemic apoptosis in the hippocampus of mice deficient in interleukin-1

The cytokine interleukin‐1 (IL‐1) has been implicated in ischemic brain damage, because the IL‐1 receptor antagonist markedly inhibits experimentally induced neuronal loss. However, to date, no studies have demonstrated the involvement of endogenous IL‐1α and IL‐ 1β in neurodegeneration. We report here, for the first time, that mice lacking IL‐1α/β (double knockout) exhibit markedly reduced neuronal loss and apoptotic cell death when exposed to transient cardiac arrest. Furthermore, we show that, despite the reduced neuronal loss, phosphorylation of JNK/SAPK (c‐Jun NH2‐ terminal protein kinase/stress activated protein kinase) and p38 enzymes remain elevated in IL‐1 knockout mice. In contrast, the inducible nitric oxide (iNOS) immunoreactivity after global ischemia was reduced in IL‐1 knockout mice as compared with wild‐type mice. The levels of nitrite (NO2−) and nitrate (NO3−) in the hippocampus of wild‐type mice were increased with time after ischemia‐reperfusion, whereas the increase was significantly inhibited in IL‐1 knockout mice. These observations strongly suggest that endogenous IL‐1 contributes to ischemic brain damage, and this influence may act through the release of nitric oxide by iNOS. J. Comp. Neurol. 448:203–216, 2002.

Pituitary adenylate cyclase-activating polypeptide (PACAP) prevents hippocampal neurons from apoptosis by inhibiting JNK/SAPK and p38 signal transduction pathways

We have demonstrated that ischemic neuronal death (apoptosis) of rat CA1 region of the hippocampus was prevented by infusing pituitary adenylate cyclase-activating polypeptide (PACAP) either intracerebroventricularly or intravenously. We have also demonstrated that the activity of mitogen-activated protein (MAP) kinase family members, including ERK (extracellular signal-regulated kinase), Jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) and p38, was increased in the hippocampus within 1-6 h after brain ischemia. The molecular mechanisms underlying the PACAP anti-apoptotic effect were demonstrated in this study. Ischemic stress had a strong influence on MAP kinase family, especially on JNK/SAPK and p38. PACAP inhibited the activation of JNK/SAPK and p38 after ischemic stress, while ERK is not suppressed. These findings suggest that PACAP inhibits the JNK/SAPK and p38 signaling pathways, thereby protecting neurons against apoptosis.

Morphological evidence for neural interactions between leptin and orexin in the hypothalamus.

Both leptin and orexin have been recently discovered as peptides involved in feeding regulation. The morphological evidence of neural interaction between leptin and orexin, one considered to inhibit food intake and the other to stimulate it in the central nervous system (CNS), was studied by use of double immunostaining method. The leptin receptor-like immunoreactive (LR-LI) neurons in the hypothalamic arcuate nucleus and ventromedial nucleus were innervated by orexin-like immunoreactive (OX-LI) neurons. The distribution of LR-LI neurons in the hypothalamus was very similar to that of OX-LI neurons. These results may suggest that leptin and orexin are intimately correlated with each other and that they reciprocally regulate feeding at the hypothalamic level.

Aneurysm of the distal anterior inferior cerebellar artery at the medial branch: a case report and review of the literature

BACKGROUND Distal aneurysms of the anterior inferior cerebellar artery (AICA) are rare. Most have been located at the cerebellopontine angle. Only three cases, including ours, located in the medial branch of the AICA have been reported in the literature. METHODS A 55-year-old female presented with epilepsy that she experienced for the first time. Computed tomography and magnetic resonance imaging revealed no abnormality. Digital subtraction angiography (DSA) disclosed a rare aneurysm at the medial branch of the distal anterior inferior cerebellar artery. In the blood workup on her admission, a mild inflammatory sign was found, and bacterial aneurysm was suggested as the presumptive differential diagnosis. Repeated DSA was performed 3 months after admission, but the aneurysm did not disappear. She then underwent a suboccipital craniotomy in the prone position. RESULTS The aneurysm was clipped and partially resected for the pathological examination. Histologic examination revealed a true aneurysm. She was discharged without neurological deficit. CONCLUSIONS In this report, we summarize the previous cases and discuss the location and clinical manifestations of aneurysms of the AICA through a review of the literature.

Regional differences in PACAP transport across the blood–brain barrier in mice: a possible influence of strain, amyloid β protein, and age

The blood-brain barrier (BBB) controls the exchange of peptides and regulatory proteins between the central nervous system (CNS) and the blood. Transport across the BBB of such regulatory substances is altered in animal models of Alzheimers disease. These alterations could lead to cognitive impairments or diminish their therapeutic potential. Here, we measured the transport rate of radioactively labeled pituitary adenylate cyclase-activating polypeptide (PACAP) from blood into whole brain and into 11 brain regions in three groups of mice: young (2 months old) ICR, young (2 months old) SAMP8, and aged (12 months old) SAMP8 mice. The SAMP8 is a strain which develops impaired learning and memory with aging that correlates with an age-related increase in brain levels of amyloid beta protein (A beta P). PACAP is a powerful neurotrophin that may have a therapeutic role in neurodegenerative diseases. We found that I-PACAP crossed the BBB fastest at the hypothalamus and the hippocampus in all three groups. Slower transport rates into the whole brain, the olfactory bulb, the hypothalamus, and the hippocampus for aged SAMP8 mice was likely related to differences both from strain and expression of A beta P with aging.

PACAP activates PKA, PKC and Ca2+ signaling cascades in rat neuroepithelial cells

Several studies have reported that the PAC(1) receptor (PAC1-R), the specific receptor for PACAP, is expressed at early developmental stages. Here, we describe that the cytosolic Ca(2+) concentration ([Ca(2+)](i)) was increased by PACAP, but not VIP, in a concentration range from 10(-12) to 10(-8) M via the PAC(1)-R in isolated single cells from the rat neural fold. This activation of the cells by PACAP was mimicked by agonists and inhibited by antagonists of the cAMP/PKA and PLC/PKC cascades. These data indicate that PACAP/PAC(1)-R is linked to [Ca(2+)](i) signaling via two G-protein-coupled protein kinase pathways and may thereby play an important role in early neurodevelopment.

Midbrain hemorrhage presenting with oculomotor nerve palsy: case report

BACKGROUND We report a case of isolated oculomotor nerve palsy caused by a midbrain hemorrhage. CASE DESCRIPTION A 75-year-old man visited our hospital complaining of double vision and left eye ptosis without headache. Neuro-ophthalmic examination showed that his left and right pupils were 3.5 mm and 3 mm in diameter, respectively, that left downward eye movement was limited, and that convergence of the right eye was limited. Magnetic resonance imaging (MRI) demonstrated that there was a hematoma located in the anterior tegmentum of the left midbrain. Two weeks after admission and treatment, including conservative therapy, his double vision gradually disappeared. CONCLUSION To date, 73 cases have been reported in the literature. Most cases of isolated oculomotor nerve palsy have been caused by diabetes mellitus, aneurysm, or infarction. However, focal midbrain hemorrhage incidentally produces third nerve palsy. MRI is extremely helpful in diagnosing a small hemorrhage of the midbrain in such cases.

The effect of cardiac arrest on the permeability of the mouse blood-brain and blood-spinal cord barrier to pituitary adenylate cyclase activating polypeptide (PACAP)☆

Time-dependent changes in peptide transport system (PTS-6), which transports the 38 amino acid pituitary adenylate cyclase activating polypeptide (PACAP) across the blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB), were studied in mice in a cardiac arrest model. The permeability of the BSCB to radioactivity labeled I131 showed a reversible increase on Day 2-(24 h) after cardiac arrest. The BBB showed no such increase. The increase in BSCB permeability was primarily located within the thoracic region of the spinal cord. We conclude that the ischemia occurring with cardiac arrest results in a transient increase in PTS-6 activity located primarily in the thoracic region of the spinal cord.