- Email: [email protected]
Ultrastructural evidence for apoptotic neuronal changes in frontal cortex of alzheimer's disease
Poster A. Clinical
s12
mediating behavioral depression (Weiss and Simson, 1985). With the potassium permanganate (KMnO,) method for electron microscopy (EM), designed to demonstrate catecholamines as dense cores inside vesicles (Hokfelt and Jonsson, 1968), we have shown that in the normal human LC perikarya numerous large globules contain dense cores, presumably indicating the presence of NA (Issidorides et al., 1996). This is a new phenotype, compared to animals, where few and small dense core vesicles are usually present in these neurons. This accumulation of large dense core globules in man could represent a back-up storage compartment of neurotransmitter necessary for meeting the increased demands to cope with unpredictable stresses, inherent in man’s ecological (social) environment. Our objective was to confirm the involvement of the globules in the fall of NA in depression by studying ten suicides with a diagnosis of major depression. We applied the KMnO,/EM method, as well as a rabbit monoclonal anti-serum to NA, visualized by colloidal gold EM immunostaining. We found that the globules in the LC neurons of the suicide cases were as numerous as those in normal controls, but their electron density was greatly reduced, indicating depletion of NA. This was confirmed by the decreased immunoreactivity of NA. Abnormalities in the ultrastructure of the double membranes surrounding the dense bodies, such as dissociations, splits and blebs, indicating increased membrane fluidity, support the hypothesis that defective membrane chemistry and structure may be the cause of this neurotransmitter leakage. Membrane fluidization, a result of low cholesterol, could be an important biological substratum in major depression for the wide spectrum of dysfunctions which are expressed in many body systems aside from the brain.
References I. Weiss, J.M., Simson, P.G., (1985) Neurochemical basis of stressinduced depression. Psychopharmacol. Bull. 21, 447-457. 2. Hokfelt, T., Jonsson, G., (1968) Studies on reaction and binding of monoamines after fixation and processing for electron microscopy with special reference to fixation with potassium permanganate. Histochemie 16, 45-67. 3. Issidorides, M.R., Kriho,V., Pappas, G.D., (1996) The fine structure of large dense-core organelles in human locus coeruleus neurons. Neurol. Res. 18, 57-63.
References Smale, G., Nichols, N.R., Brady, D.R., Finch, C.E., Horton, W.E., Jr (1995) Evidence for apoptotic cell death in Alzheimer’s disease. Exp. Neurol. 133, 225-230. Su, J.H., Anderson, A.J., Cummings, B.J., Cotman, C.W., (1994) hnmunohistochemical evidence for apoptosis in Alzheimer’s disease. Neuroreport 5, 2529-2533. Issidorides, M.R., Katsorchis, T., (1981) Dispersed and compact chromatin demonstrated with a new EM method: Phosphotungstic acid hematoxylin block-staining. Histochemistry 73, 21-31.
A-13 El
Brain perfusion spet in unipolar major depression before and after treatment
Margareta Kocmur”, Metka Mil&nskih, Natasa V Budihna’. “Psychiatry, ‘Nuclear Medicine, University Medical Centre, ‘Nuclear Medicine, Oncology, 1525 .Ljubljana, Slovenin Aim of our study was to evaluate regional cerebral blood flow (r-CBF) in unipolar depression before therapy and the effect of antidepressant drugs on r-CBF. Methods: Clinical criteria for depression without psychosis were met according to psychiatric evaluation. Severity of depression was assessed with Hamilton Depression Rating Scale (HAMD) just before every scintigraphic study. r-CBF was measured using 99m-Tc-HMPAO-SPET or 99m-Tc-ECD in 10 patients with severe unipolar depression before beginning of the antidepressant drug therapy, three weeks and 6 months after. Only pts with no change in antidepressant medication during the study were included. No antipsychotic drug was used. In each patient r-CBF was compared to cerebellar blood flow in all studies and separate regions were evaluated on repeated studies. r-CBF was evaluated semiquantitatively as equal to cerebellum, decreased, and severely decreased. CBF in basal ganglia vs. cortex was scored as equal, diminished or increased. Results: all pts were clinically depressed.
This work was supported by the Theodor Theohari Cozzika Foundation and the Ministry of Development, General Secretariat of Research and Technology (Grant No 90Hz124). Greece b&xc
ptr
HAMD
r-CBF
r-CBF
number
average
fmnlal
temporal
parietal
range
awage
average
average
25.3
19
2.1
?I
2.4
2.6
24
2.6
29
28
IO
therapy
A-12 El
Ultrastructural evidence for apoptotic neuronal changes in frontal cortex of alzheimer’s disease
M.R. lssidorides”.b M. Chrysanthou-Piterou”, S. Havaki’, V. Krihob and G.D. Pappasb. “Department of Psychiatry, University of Athens and ‘Department of Anatomy & Cell Biology, University of Illinois at Chicago, USA Neuronal death is a prominent feature of Alzheimer’s disease (AD). Evidence has been provided, using the in situ labeling technique TUNEL, for apoptosis occurring in a high incidence in the hippocampus of AD patients compared to controls (Smale et al., 1995). The main morphological characteristics of apoptosis are changes in the nucleus involving chromatin condensation and aggregation into dense, often crescent-shaped masses under the nuclear membrane and around the nucleolus (Su et al., 1994). We designed to study apoptosis in the frontal cortex of 3 AD patients and 3 controls, using the phosphotungstic acid hematoxylin (PTAH) in to the staining method for electron microscopy (EM) after glutaraldehyde fixation. PTAH differentiates loose from condensed chromatin in a “black and white” contrast (Issidorides and Katsorchis, 198 I), so that results are evaluated by reliable ultrastructural morphological criteria. In the patients’ samples, we found several neurons in various stages of chromatin condensation and nuclear fragmentation, which are similar to the stages occurring in apoptotic human epidermal keratinocytes that usually precede the cleavage of DNA. These EM data offer the possibility to pin-point the initial site of chromatin condensation, i.e. the site of origin of the apoptotic process, and probe us to search for inhibitory factors and processes targeted to the prevention of its evolution. (Supported by NATO Collaborative Research Grant No 910753 and by Grant No 90HZl24 of the General Secretariat of Research and Technology, Greece)
3 week\
r-CBF
16-32 Y
157 9-23
6 months
s
6.5 6-7
Increase in perfusion was highly significant (p
IA-14
Serotonin content in blood plasma and in platelets of patlents with panic disorder
B.M. Kogan, AZ. Drozdov, E.V. Koroleva, E.D. Jukovsky. Serbsky National Research Centre for Social and Forensic Psychiatv, Moscow, Russia A lot of evidence suggests that serotonin system plays a significant role in the pathogenesis of affective disorders. For example, the effectiveness of serotoliin reuptake inhibitors in treating depressive, obsessive-compulsive, anxiety, and panic disorder has been demonstrated. However, the precise role of serotonergic system in the pathogenesis of panic attacks is still unclear. Particularly, there are some contradictory results in regard to level of sexotonin uptake in human platelets in panic disorder.
s12
mediating behavioral depression (Weiss and Simson, 1985). With the potassium permanganate (KMnO,) method for electron microscopy (EM), designed to demonstrate catecholamines as dense cores inside vesicles (Hokfelt and Jonsson, 1968), we have shown that in the normal human LC perikarya numerous large globules contain dense cores, presumably indicating the presence of NA (Issidorides et al., 1996). This is a new phenotype, compared to animals, where few and small dense core vesicles are usually present in these neurons. This accumulation of large dense core globules in man could represent a back-up storage compartment of neurotransmitter necessary for meeting the increased demands to cope with unpredictable stresses, inherent in man’s ecological (social) environment. Our objective was to confirm the involvement of the globules in the fall of NA in depression by studying ten suicides with a diagnosis of major depression. We applied the KMnO,/EM method, as well as a rabbit monoclonal anti-serum to NA, visualized by colloidal gold EM immunostaining. We found that the globules in the LC neurons of the suicide cases were as numerous as those in normal controls, but their electron density was greatly reduced, indicating depletion of NA. This was confirmed by the decreased immunoreactivity of NA. Abnormalities in the ultrastructure of the double membranes surrounding the dense bodies, such as dissociations, splits and blebs, indicating increased membrane fluidity, support the hypothesis that defective membrane chemistry and structure may be the cause of this neurotransmitter leakage. Membrane fluidization, a result of low cholesterol, could be an important biological substratum in major depression for the wide spectrum of dysfunctions which are expressed in many body systems aside from the brain.
References I. Weiss, J.M., Simson, P.G., (1985) Neurochemical basis of stressinduced depression. Psychopharmacol. Bull. 21, 447-457. 2. Hokfelt, T., Jonsson, G., (1968) Studies on reaction and binding of monoamines after fixation and processing for electron microscopy with special reference to fixation with potassium permanganate. Histochemie 16, 45-67. 3. Issidorides, M.R., Kriho,V., Pappas, G.D., (1996) The fine structure of large dense-core organelles in human locus coeruleus neurons. Neurol. Res. 18, 57-63.
References Smale, G., Nichols, N.R., Brady, D.R., Finch, C.E., Horton, W.E., Jr (1995) Evidence for apoptotic cell death in Alzheimer’s disease. Exp. Neurol. 133, 225-230. Su, J.H., Anderson, A.J., Cummings, B.J., Cotman, C.W., (1994) hnmunohistochemical evidence for apoptosis in Alzheimer’s disease. Neuroreport 5, 2529-2533. Issidorides, M.R., Katsorchis, T., (1981) Dispersed and compact chromatin demonstrated with a new EM method: Phosphotungstic acid hematoxylin block-staining. Histochemistry 73, 21-31.
A-13 El
Brain perfusion spet in unipolar major depression before and after treatment
Margareta Kocmur”, Metka Mil&nskih, Natasa V Budihna’. “Psychiatry, ‘Nuclear Medicine, University Medical Centre, ‘Nuclear Medicine, Oncology, 1525 .Ljubljana, Slovenin Aim of our study was to evaluate regional cerebral blood flow (r-CBF) in unipolar depression before therapy and the effect of antidepressant drugs on r-CBF. Methods: Clinical criteria for depression without psychosis were met according to psychiatric evaluation. Severity of depression was assessed with Hamilton Depression Rating Scale (HAMD) just before every scintigraphic study. r-CBF was measured using 99m-Tc-HMPAO-SPET or 99m-Tc-ECD in 10 patients with severe unipolar depression before beginning of the antidepressant drug therapy, three weeks and 6 months after. Only pts with no change in antidepressant medication during the study were included. No antipsychotic drug was used. In each patient r-CBF was compared to cerebellar blood flow in all studies and separate regions were evaluated on repeated studies. r-CBF was evaluated semiquantitatively as equal to cerebellum, decreased, and severely decreased. CBF in basal ganglia vs. cortex was scored as equal, diminished or increased. Results: all pts were clinically depressed.
This work was supported by the Theodor Theohari Cozzika Foundation and the Ministry of Development, General Secretariat of Research and Technology (Grant No 90Hz124). Greece b&xc
ptr
HAMD
r-CBF
r-CBF
number
average
fmnlal
temporal
parietal
range
awage
average
average
25.3
19
2.1
?I
2.4
2.6
24
2.6
29
28
IO
therapy
A-12 El
Ultrastructural evidence for apoptotic neuronal changes in frontal cortex of alzheimer’s disease
M.R. lssidorides”.b M. Chrysanthou-Piterou”, S. Havaki’, V. Krihob and G.D. Pappasb. “Department of Psychiatry, University of Athens and ‘Department of Anatomy & Cell Biology, University of Illinois at Chicago, USA Neuronal death is a prominent feature of Alzheimer’s disease (AD). Evidence has been provided, using the in situ labeling technique TUNEL, for apoptosis occurring in a high incidence in the hippocampus of AD patients compared to controls (Smale et al., 1995). The main morphological characteristics of apoptosis are changes in the nucleus involving chromatin condensation and aggregation into dense, often crescent-shaped masses under the nuclear membrane and around the nucleolus (Su et al., 1994). We designed to study apoptosis in the frontal cortex of 3 AD patients and 3 controls, using the phosphotungstic acid hematoxylin (PTAH) in to the staining method for electron microscopy (EM) after glutaraldehyde fixation. PTAH differentiates loose from condensed chromatin in a “black and white” contrast (Issidorides and Katsorchis, 198 I), so that results are evaluated by reliable ultrastructural morphological criteria. In the patients’ samples, we found several neurons in various stages of chromatin condensation and nuclear fragmentation, which are similar to the stages occurring in apoptotic human epidermal keratinocytes that usually precede the cleavage of DNA. These EM data offer the possibility to pin-point the initial site of chromatin condensation, i.e. the site of origin of the apoptotic process, and probe us to search for inhibitory factors and processes targeted to the prevention of its evolution. (Supported by NATO Collaborative Research Grant No 910753 and by Grant No 90HZl24 of the General Secretariat of Research and Technology, Greece)
3 week\
r-CBF
16-32 Y
157 9-23
6 months
s
6.5 6-7
Increase in perfusion was highly significant (p
IA-14
Serotonin content in blood plasma and in platelets of patlents with panic disorder
B.M. Kogan, AZ. Drozdov, E.V. Koroleva, E.D. Jukovsky. Serbsky National Research Centre for Social and Forensic Psychiatv, Moscow, Russia A lot of evidence suggests that serotonin system plays a significant role in the pathogenesis of affective disorders. For example, the effectiveness of serotoliin reuptake inhibitors in treating depressive, obsessive-compulsive, anxiety, and panic disorder has been demonstrated. However, the precise role of serotonergic system in the pathogenesis of panic attacks is still unclear. Particularly, there are some contradictory results in regard to level of sexotonin uptake in human platelets in panic disorder.