Expression of the yes proto-oncogene in cerebellar Purkinje cells.

To identify the kinds of cells in the brain that express the yes proto-oncogene, we examined chicken brains by using immunofluorescent staining and in situ hybridization. Both approaches showed that the highest level of the yes gene product was in cerebellar Purkinje cells. In addition, we analyzed Purkinje cell degeneration (pcd) mutant mice. The level of yes mRNA in cerebella of pcd mutants was four times lower than that found in cerebella of normal littermates. Our studies point to Purkinje cells as an attractive model for functional studies of the yes protein.

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Cellular distribution of the RNA transcripts of a newly discovered gene in the brain of normal, weaver, Purkinje cell degeneration and reeler mutant mice as evidenced by in situ hybridization histochemistry

Molecular Brain Research ◽

10.1016/0169-328x(93)90096-8 ◽

1993 ◽

Vol 18 (4) ◽

pp. 321-328 ◽

Cited By ~ 5

Author(s):

M. Kambouris ◽

L. Sangameswaran ◽

S.R. Dlouhy ◽

M.E. Hodes ◽

B. Ghetti ◽

...

Keyword(s):

In Situ Hybridization ◽

Purkinje Cell ◽

Cellular Distribution ◽

Cell Degeneration ◽

Hybridization Histochemistry ◽

Rna Transcripts ◽

In Situ Hybridization Histochemistry ◽

Reeler Mutant ◽

The Brain

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Activation of MAP3K DLK and LZK in Purkinje Cells Causes Rapid and Slow Degeneration Depending on Signaling Strength

10.1101/2020.10.09.332940 ◽

2020 ◽

Author(s):

Yunbo Li ◽

Erin M Ritchie ◽

Christopher L. Steinke ◽

Cai Qi ◽

Lizhen Chen ◽

...

Keyword(s):

Alternative Splicing ◽

Purkinje Cell ◽

Purkinje Cells ◽

Leucine Zipper ◽

Activity Levels ◽

Cell Type ◽

Cell Degeneration ◽

Mechanistic Basis ◽

Cerebellar Purkinje Cells ◽

Jnk Activation

SummaryThe conserved MAP3K Dual leucine zipper kinases can activate JNK via MKK4 or MKK7. Vertebrate DLK and LZK share similar biochemical activities and undergo auto-activation upon increased expression. Depending on cell-type and nature of insults DLK and LZK can induce pro-regenerative, pro-apoptotic or pro-degenerative responses, although the mechanistic basis of their action is not well understood. Here, we investigated these two MAP3Ks in cerebellar Purkinje cells using loss- and gain-of function mouse models. While loss of each or both kinases does not cause discernible defects in Purkinje cells, activating DLK causes rapid death and activating LZK leads to slow degeneration. Each kinase induces JNK activation and caspase-mediated apoptosis independent of each other. Significantly, deleting CELF2, which regulates alternative splicing of Mkk7, strongly attenuates Purkinje cell degeneration induced by activation of LZK, but not DLK. Thus, controlling the activity levels of DLK and LZK is critical for neuronal survival and health.

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Disrupted Calcium Signaling in Animal Models of Human Spinocerebellar Ataxia (SCA)

International Journal of Molecular Sciences ◽

10.3390/ijms21010216 ◽

2019 ◽

Vol 21 (1) ◽

pp. 216 ◽

Cited By ~ 3

Author(s):

Francesca Prestori ◽

Francesco Moccia ◽

Egidio D’Angelo

Keyword(s):

Animal Models ◽

Purkinje Cell ◽

Calcium Signaling ◽

Purkinje Cells ◽

Motor Coordination ◽

Cerebellar Atrophy ◽

Cerebellar Nuclei ◽

Spinocerebellar Ataxias ◽

Cell Degeneration ◽

Cerebellar Purkinje Cells

Spinocerebellar ataxias (SCAs) constitute a heterogeneous group of more than 40 autosomal-dominant genetic and neurodegenerative diseases characterized by loss of balance and motor coordination due to dysfunction of the cerebellum and its efferent connections. Despite a well-described clinical and pathological phenotype, the molecular and cellular events that underlie neurodegeneration are still poorly undaerstood. Emerging research suggests that mutations in SCA genes cause disruptions in multiple cellular pathways but the characteristic SCA pathogenesis does not begin until calcium signaling pathways are disrupted in cerebellar Purkinje cells. Ca2+ signaling in Purkinje cells is important for normal cellular function as these neurons express a variety of Ca2+ channels, Ca2+-dependent kinases and phosphatases, and Ca2+-binding proteins to tightly maintain Ca2+ homeostasis and regulate physiological Ca2+-dependent processes. Abnormal Ca2+ levels can activate toxic cascades leading to characteristic death of Purkinje cells, cerebellar atrophy, and ataxia that occur in many SCAs. The output of the cerebellar cortex is conveyed to the deep cerebellar nuclei (DCN) by Purkinje cells via inhibitory signals; thus, Purkinje cell dysfunction or degeneration would partially or completely impair the cerebellar output in SCAs. In the absence of the inhibitory signal emanating from Purkinje cells, DCN will become more excitable, thereby affecting the motor areas receiving DCN input and resulting in uncoordinated movements. An outstanding advantage in studying the pathogenesis of SCAs is represented by the availability of a large number of animal models which mimic the phenotype observed in humans. By mainly focusing on mouse models displaying mutations or deletions in genes which encode for Ca2+ signaling-related proteins, in this review we will discuss the several pathogenic mechanisms related to deranged Ca2+ homeostasis that leads to significant Purkinje cell degeneration and dysfunction.

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Activation of MAP3K DLK and LZK in Purkinje cells causes rapid and slow degeneration depending on signaling strength

eLife ◽

10.7554/elife.63509 ◽

2021 ◽

Vol 10 ◽

Author(s):

Yunbo Li ◽

Erin M Ritchie ◽

Christopher L Steinke ◽

Cai Qi ◽

Lizhen Chen ◽

...

Keyword(s):

Alternative Splicing ◽

Purkinje Cell ◽

Purkinje Cells ◽

Leucine Zipper ◽

Activity Levels ◽

Cell Type ◽

Cell Degeneration ◽

Mechanistic Basis ◽

Cerebellar Purkinje Cells ◽

Jnk Activation

The conserved MAP3K Dual-Leucine-Zipper Kinase (DLK) and Leucine-Zipper-bearing Kinase (LZK) can activate JNK via MKK4 or MKK7. These two MAP3Ks share similar biochemical activities and undergo auto-activation upon increased expression. Depending on cell-type and nature of insults DLK and LZK can induce pro-regenerative, pro-apoptotic or pro-degenerative responses, although the mechanistic basis of their action is not well understood. Here, we investigated these two MAP3Ks in cerebellar Purkinje cells using loss- and gain-of function mouse models. While loss of each or both kinases does not cause discernible defects in Purkinje cells, activating DLK causes rapid death and activating LZK leads to slow degeneration. Each kinase induces JNK activation and caspase-mediated apoptosis independent of each other. Significantly, deleting CELF2, which regulates alternative splicing of Map2k7, strongly attenuates Purkinje cell degeneration induced by LZK, but not DLK. Thus, controlling the activity levels of DLK and LZK is critical for neuronal survival and health.

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Purkinje cell degeneration associated with erythroid ankyrin deficiency in nb/nb mice.

The Journal of Cell Biology ◽

10.1083/jcb.114.6.1233 ◽

1991 ◽

Vol 114 (6) ◽

pp. 1233-1241 ◽

Cited By ~ 68

Author(s):

L L Peters ◽

C S Birkenmeier ◽

R T Bronson ◽

R A White ◽

S E Lux ◽

...

Keyword(s):

Purkinje Cell ◽

Purkinje Cells ◽

Granule Cells ◽

Cell Loss ◽

Chromosome 8 ◽

Cell Degeneration ◽

Purkinje Cell Loss ◽

Cell Stability ◽

Cerebellar Purkinje Cells

Mice homozygous for the nb mutation (Chromosome 8) have a severe hemolytic anemia and develop a psychomotor disorder at 6 mo of age. The nb/nb mice are deficient in erythroid ankyrin (Ank-1) but, until the present study, the role of Ank-1 and of Ank-2 (brain ankyrin) in disease genesis was unknown. In normal erythroid tissues, we show that two major transcripts are expressed from Ank-1, and one of these is also present at high levels in the cerebellum. By in situ hybridization and immunocytochemistry, Ank-1 localizes to the cerebellar Purkinje cells and, to a lesser extent, the granule cells. In nb/nb mice, Ank-1 transcripts are markedly reduced in both erythroid and neural tissue, and nb/nb Purkinje cells and granule cells are nearly devoid of Ank-1. The neurological syndrome appears concurrently with a dramatic loss of Purkinje cells. Ank-2 maps to Chromosome 3 and its expression is unaffected by the nb mutation. We conclude that Ank-1 is specifically required for Purkinje cell stability and, in its absence, Purkinje cell loss and neurological symptoms appear.

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Glucose transporter expression in brain. cDNA sequence of mouse GLUT3, the brain facilitative glucose transporter isoform, and identification of sites of expression by in situ hybridization.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)48518-3 ◽

1992 ◽

Vol 267 (1) ◽

pp. 467-472

Author(s):

S Nagamatsu ◽

J M Kornhauser ◽

C F Burant ◽

S Seino ◽

K E Mayo ◽

...

Keyword(s):

In Situ Hybridization ◽

Cdna Sequence ◽

Glucose Transporter ◽

The Brain ◽

Transporter Expression ◽

Facilitative Glucose Transporter

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The lurcher gene induces apoptotic death in cerebellar Purkinje cells

Development ◽

10.1242/dev.121.4.1183 ◽

1995 ◽

Vol 121 (4) ◽

pp. 1183-1193 ◽

Cited By ~ 1

Author(s):

D.J. Norman ◽

L. Feng ◽

S.S. Cheng ◽

J. Gubbay ◽

E. Chan ◽

...

Keyword(s):

Purkinje Cells ◽

Nuclear Dna ◽

Neuronal Loss ◽

Terminal Phase ◽

Nuclear Condensation ◽

Electron Microscopic ◽

Apoptotic Death ◽

Microscopic Studies ◽

Cerebellar Purkinje Cells

In the neurologically mutant mouse strain lurcher (Lc), heterozygous animals display cell autonomous degeneration of cerebellar Purkinje cells beginning in the second postnatal week. During the course of our studies to identify the genetic lesion responsible for this disease (Norman et al., 1991), we have formulated an hypothesis suggesting that in Lc Purkinje cells homeostasis is sufficiently perturbed to lead to the activation of programmed cell death, thus resulting in neuronal loss and the consequent neurologic disease (Heintz, 1993). To address this possibility, we have examined the properties of Lc Purkinje cells as they die during the second postnatal week. Our light and electron microscopic studies demonstrate that dying Lc Purkinje cells exhibit the characteristic morphologic features of apoptosis, including nuclear condensation, axon beading and membrane blebbing. Using an in situ end-labeling method, we have also detected nicked nuclear DNA in these cells. Furthermore, we have examined the expression of the sulfated glycoprotein 2 (SGP2), whose mRNA is induced in both T-cells and prostate epithelial cells undergoing apoptotic death. We show by in situ hybridization that SGP2 is not expressed at detectable levels in normal Purkinje cells, but that its mRNA is present in Lc Purkinje cells prior to their death. Also expression of the Kv3.3b potassium channel, which marks the terminal phase of Purkinje cell differentiation, is evident in Lc Purkinje cells prior to their death. These data demonstrate that the Lc mutation induces apoptosis in cerebellar Purkinje cells following their maturation in postnatal cerebellum. Isolation of the Lc mutation and further analysis of its action in eliciting apoptosis can provide an important opportunity for understanding the etiology of neurodegenerative disease.

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Expression of vascular permeability factor/vascular endothelial growth factor in normal rat tissues

AJP Cell Physiology ◽

10.1152/ajpcell.1993.264.4.c995 ◽

1993 ◽

Vol 264 (4) ◽

pp. C995-C1002 ◽

Cited By ~ 240

Author(s):

W. T. Monacci ◽

M. J. Merrill ◽

E. H. Oldfield

Keyword(s):

Vascular Endothelial Growth Factor ◽

In Situ Hybridization ◽

Rat Tissues ◽

Vascular Endothelial ◽

Vascular Permeability Factor ◽

Organ Specific ◽

Normal Rat ◽

Endothelial Growth Factor ◽

The Brain

Vascular permeability factor (VPF)/vascular endothelial growth factor (VEGF) is a approximately 43-kDa secreted protein that has been shown in bioassays to induce endothelial proliferation, angiogenesis, and capillary hyperpermeability. VPF has been suggested to play an important role in the physiology of normal vasculature. To further elucidate the natural functions of VPF in vivo, the expression of VPF in normal tissues was examined using Northern blot analysis and in situ hybridization histochemistry. VPF mRNA is expressed in the brain, kidney, liver, lung, and spleen of the healthy adult rat. On Northern blots, the relative abundance of VPF mRNA observed in these tissues was highest in the lung and lowest in the spleen. As determined by in situ hybridization, the patterns of VPF expression are organ specific. Hybridization of an antisense VPF probe was concentrated in the cerebellar granule cell layer of the brain and in the glomeruli and tubules of the kidney. In the liver and lung, intense hybridization was observed homogeneously throughout both tissues, demonstrating that VPF mRNA is present in virtually every hepatocyte and pulmonary alveolar cell. Hybridization to the spleen was weaker and more diffuse. The widespread expression and organ-specific distribution of VPF mRNA in normal rat tissues supports the suggestion of an extensive role for this factor in the physiology of normal vasculature.

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