Targeting cell death in vivo in experimental traumatic brain injury by a novel molecular probe.
Traumatic brain injury (TBI) remains a frequent and major challenge in neurological and neurosurgical practice. Apoptosis may play a role in cerebral tissue damage induced by the traumatic insult, and thus its detection and inhibition may advance patient care. DDC (N,N'-didansyl-L-cystine) is a novel fluorescent probe for detection of apoptotic cells. We now report on the performance of DDC in experimental TBI. Closed head injury was induced in mice by weight-drop. DDC was administered intravenously in vivo. Two hours later, animals were sacrificed, and brain tissue was subjected to fluorescent microcopy, for assessment of DDC uptake, in correlation with histopathological assessment of apoptosis by TUNEL and caspase substrates, and also in correlation with the neurological deficits, as assessed by Neurological Severity Score (NSS). Selective uptake of DDC was observed at the primary site of injury, and also at remote sites. Uptake was at the cellular level, with accumulation of DDC in the cytoplasm. Cells manifesting DDC uptake were confirmed as apoptotic cells by detection of the characteristic apoptotic DNA fragmentation (positive TUNEL staining) and detection of activated caspases. The damaged region stained by DDC fluorescence correlated with the severity of neuronal deficits. Our study confirms the role of apoptosis in TBI, and proposes DDC as a useful tool for its selective targeting and detection in vivo. Such imaging of apoptosis, following future radiolabeling of DDC, may advance care for patients with head injury, by allowing real-time evaluation of the extent of tissue damage, assessment of novel therapeutic strategies, and optimization of treatment for the individual patient.
Reshef
Ayelet
A
NST NeuroSurvival Technologies Ltd., Petach-Tikva, Israel.
Shirvan
Anat
A
Shohami
Esther
E
Grimberg
Hagit
H
Levin
Galit
G
Cohen
Avi
A
Trembovler
Victoria
V
Ziv
Ilan
I
D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001921:Brain / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D001930:Brain Injuries / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D020169:Caspases / Q000378:metabolism; D003553:Cystine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism / Q000473:pathology; D053938:DNA Fragmentation; D003619:Dansyl Compounds / Q000737:chemistry / Q000378:metabolism; D004195:Disease Models, Animal; D018450:Disease Progression; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D051379:Mice; D008807:Mice, Inbred BALB C; D015394:Molecular Structure; D009410:Nerve Degeneration / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D011237:Predictive Value of Tests; D013194:Staining and Labeling / Q000379:methods
D051017:Apoptosis Regulatory Proteins; D003619:Dansyl Compounds; D005456:Fluorescent Dyes; C038494:di(dansyl)cystine; D003553:Cystine; D020169:Caspases