Abstract
Human brain samples were collected from 70 autopsy cases including 22 freshwater drowning (FWD), 26 saltwater drowning (SWD), and 22 non-drowning cases as controls. Then, immunohistochemical study combined with morphometry was carried out in order to examine the differential expression of AQP1 and AQP4 in the brain samples. Immunohistochemically, star-shaped cells bearing highly branched processes, often surrounding blood vessels, showed positive reactions for AQP1 and AQP4 in FWD, SWD, as well as control groups. Additionally, with double-color immunofluorescence analysis, AQP1- or AQP4-positive cells could be identified as GFAP-positive astrocytes. Moreover, AQP1-positive reaction was also observed in blood vessels. Morphometrically, there were no significant differences in AQP1 expression in astrocytes or in blood vessels among the three groups. In contrast, the average value of AQP4-positive astrocytes was significantly higher in FWD cases than in SWD and control groups. Moreover, AQP4 expression was significantly lower in SWD than in the control group (p < 0.05). Moreover, there was no significant correlation between post-submerged interval and AQP expression in drowning cases. Therefore, immunohistochemical analysis of intracerebral AQP4 expression would be forensically useful for differentiation between FWD and SWD.
References
Piette MH, De Letter EA (2006) Drowning: still a difficult autopsy diagnosis. Forensic Sci Int 163:1–9
Hurlimann J, Feer P, Elber F, Niederberger K, Dirnhofer R, Wyler D (2000) Diatom detection in the diagnosis of death by drowning. Int J Leg Med 114:6–14
Farrugia A, Ludes B (2010) Diatomeennachweis und –identifizierung. Rechtsmedizin 20:49–58
Brinkmann B, Fechner G, Puschel K (1983) Ultrastructural pathology of the alveolar apparatus in experimental drowning. Z Rechtsmed 91:47–60
Brinkmann B, Fechner G, Puschel K (1983) Lung histology in experimental drowning. Z Rechtsmed 89:267–277
Zhu BL, Quan L, Li DR, Taniguchi M, Kamikodai Y, Tsuda K, Fujita MQ, Nishi K, Tsuji T, Maeda H (2003) Postmortem lung weight in drownings: a comparison with acute asphyxiation and cardiac death. Leg Med (Tokyo) 5:20–26
de la Grandmaison GL, Leterreux M, Lasseuguette K, Alvarez JC, de Mazancourt P, Durigon M (2006) Study of the diagnostic value of iron in fresh water drowning. Forensic Sci Int 157:117–120
Azparren JE, Fernandez-Rodriguez A, Vallejo G (2003) Diagnosing death by drowning in fresh water using blood strontium as an indicator. Forensic Sci Int 137:55–59
Zhu BL, Ishida K, Taniguchi M, Quan L, Oritani S, Tsuda K, Kamikodai Y, Fujita MQ, Maeda H (2003) Possible postmortem serum markers for differentiation between fresh-, saltwater drowning and acute cardiac death: a preliminary investigation. Leg Med (Tokyo) 5(Suppl 1):S298–301
Kakizaki E, Kozawa S, Matsuda H, Muraoka E, Uchiyama T, Sakai M, Yukawa N (2010) Freshwater bacterioplankton cultured from liver, kidney and lungs of a decomposed cadaver retrieved from a sandy seashore: possibility of drowning in a river and then floating out to sea. Leg Med (Tokyo) 12:195–199
Nielsen S, Frokiaer J, Marples D, Kwon TH, Agre P, Knepper MA (2002) Aquaporins in the kidney: from molecules to medicine. Physiol Rev 82:205–244
Verkman AS (2005) More than just water channels: unexpected cellular roles of aquaporins. J Cell Sci 118:3225–3232
Gunnarson E, Zelenina M, Aperia A (2004) Regulation of brain aquaporins. Neuroscience 129:947–955
Vajda Z, Promeneur D, Doczi T, Sulyok E, Frokiaer J, Ottersen OP, Nielsen S (2000) Increased aquaporin-4 immunoreactivity in rat brain in response to systemic hyponatremia. Biochem Biophys Res Commun 270:495–503
Misawa T, Arima K, Mizusawa H, Satoh J (2008) Close association of water channel AQP1 with amyloid-beta deposition in Alzheimer disease brains. Acta Neuropathol 116:247–260
Tran ND, Kim S, Vincent HK, Rodriguez A, Hinton DR, Bullock MR, Young HF (2010) Aquaporin-1-mediated cerebral edema following traumatic brain injury: effects of acidosis and corticosteroid administration. J Neurosurg 112:1095–1104
Ding JY, Kreipke CW, Speirs SL, Schafer P, Schafer S, Rafols JA (2009) Hypoxia-inducible factor-1α signaling in aquaporin upregulation after traumatic brain injury. Neurosci Lett 453:68–72
Suzuki R, Okuda M, Asai J et al (2006) Astrocytes co-express aquaporin-1, -4, and vascular endothelial growth factor in brain edema tissue associated with brain contusion. Acta Neurochir Suppl 96:398–401
Satoh J, Tabunoki H, Yamamura T, Arima K, Konno H (2007) Human astrocytes express aquaporin-1 and aquaporin-4 in vitro and in vivo. Neuropathology 27:245–256
Manley GT, Fujimura M, Ma T, Noshita N, Filiz F, Bollen AW, Chan P, Verkman AS (2000) Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke. Nat Med 6:159–163
Papadopoulos MC, Manley GT, Krishna S, Verkman AS (2004) Aquaporin-4 facilitates reabsorption of excess fluid in vasogenic brain edema. FASEB J 18:1291–1293
Hayashi T, Ishida Y, Mizunuma S, Kimura A, Kondo T (2009) Differential diagnosis between freshwater drowning and saltwater drowning based on intrapulmonary aquaporin-5 expression. Int J Leg Med 123:7–13
An JL, Ishida Y, Kimura A, Kondo T (2009) Forensic application of intrarenal aquaporin-2 expression for differential diagnosis between freshwater and saltwater drowning. Int J Leg Med 124:99–104
Kitamura O (1994) Immunohistochemical investigation of hypoxic/ischemic brain damage in forensic autopsy cases. Int J Leg Med 107:69–76
Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2008) Expression of oxygen-regulated protein 150 (ORP150) in skin wound healing and its application for wound age determination. Int J Leg Med 122:409–414
Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2009) Detection of fibrocytes in human skin wounds and its application for wound age determination. Int J Leg Med 123:299–304
Zelenina M (2010) Regulation of brain aquaporins. Neurochem Int 57:468–488
Montgomery DL (1994) Astrocytes: form, functions, and roles in disease. Vet Pathol 31:145–167
Yang B, Zador Z, Verkman AS (2008) Glial cell aquaporin-4 overexpression in transgenic mice accelerates cytotoxic brain swelling. J Biol Chem 283:15280–15286
Solenov E, Watanabe H, Manley GT, Verkman AS (2004) Sevenfold-reduced osmotic water permeability in primary astrocyte cultures from AQP-4-deficient mice, measured by a fluorescence quenching method. Am J Physiol Cell Physiol 286:C426–432
Yukutake Y, Yasui M (2009) Regulation of water permeability through aquaporin-4. Neuroscience 168:885–891
Swann HG, Brucer M, Moore C (1947) Fresh water and sea water drowning; a study of the terminal cardiac and biochemical events. Tex Rep Biol Med 5:423–437
Swann HG, Brucer M (1949) The cardiorespiratory and biochemical events during rapid anoxic death; fresh water and sea water drowning. Tex Rep Biol Med 7:604–618
Zeng HK, Wang QS, Deng YY, Fang M, Chen CB, Fu YH, Jiang WQ, Jiang X (2010) Hypertonic saline ameliorates cerebral edema through downregulation of aquaporin-4 expression in the astrocytes. Neuroscience 166:878–885
Hayashi T, Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2004) Forensic application of VEGF expression to skin wound age determination. Int J Leg Med 118:320–325
Kondo T, Tanaka J, Ishida Y, Mori R, Takayasu T, Ohshima T (2002) Ubiquitin expression in skin wounds and its application to forensic wound age determination. Int J Leg Med 116:267–272
Kondo T, Ohshima T, Eisenmenger W (1999) Immunohistochemical and morphometrical study on the temporal expression of interleukin-1α (IL-1α) in human skin wounds for forensic wound age determination. Int J Leg Med 112:249–252
Kondo T, Ohshima T, Mori R, Guan DW, Ohshima K, Eisenmenger W (2002) Immunohistochemical detection of chemokines in human skin wounds and its application to wound age determination. Int J Leg Med 116:87–91
Acknowledgments
We sincerely thank Ms. Mariko Kawaguchi for her excellent assistance in preparing this manuscript. This study was financially supported in part by a Grant-in-Aid for Scientific Research (A) from the Ministry of Education, Culture, Sports, Science, and Technology of the Japanese Government.
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An, JL., Ishida, Y., Kimura, A. et al. Immunohistochemical examination of intracerebral aquaporin-4 expression and its application for differential diagnosis between freshwater and saltwater drowning. Int J Legal Med 125, 59–65 (2011). https://doi.org/10.1007/s00414-010-0523-8
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DOI: https://doi.org/10.1007/s00414-010-0523-8